Biology and Life Sciences

Abstract: Mechanotransduction is how cells convert mechanical stimuli — tension, shear, substrate stiffness, viscoelasticity and confinement — into biochemical and transcriptional signals. Once viewed as a specialty of sensory cells, it is now a general principle of cell biology, spanning scales from single-bond lifetimes to tissue remodeling and timescales from millisecond channel gating to durable epigenetic memory. This review organizes the field into five threads: adhesion and cytoskeletal force sensors; mechanosensitive ion channels; the extracellular matrix as an instructor of cell fate; nuclear mechanotransduction and chromatin remodeling; and the engineering of mechanotransduction. A final section highlights plant mechanobiology, where PIEZO channels and the receptor kinase FERONIA reveal distinct architectures relevant to membrane biophysics and biomolecular condensate biology. Throughout, force acts mainly by directly altering molecular conformation; lipids and membranes are active participants; time-dependent matrix properties often outrank static stiffness; and the genome behaves as a mechanically responsive organelle that records a cell’s mechanical history.

Abstract: Oleanolic acid (OA) is a pentacyclic triterpenoid with broad biological activity and a chemically accessible scaffold that has yielded numerous semisynthetic derivatives. Many of these compounds alter phosphorylation-dependent pathways, including PI3K/AKT/mTOR, AMPK/mTOR, MAPK, JAK/STAT3, NF-κB, and Nrf2 signaling. However, such changes are commonly reported as isolated pathway effects or down-stream responses to cellular stress, with less attention paid to the balance between kinase and phosphatase activities. This review examines OA derivatives at the kinase–phosphatase interface, with emphasis on protein tyrosine phosphatase 1B (PTP1B), re-ceptor tyrosine kinases, EGFR/AKT signaling, and phosphorylation-dependent transi-tions from stress adaptation to cancer cell death. The available evidence links OA and selected derivatives with PTP1B inhibition, receptor-proximal signaling, AKT/mTOR and AMPK/mTOR regulation, stress- and inflammation-related pathways, autophagy, and apoptosis. We also introduce the concept of structure–signaling relationships, in which chemical modification may influence not only potency, but also the point at which a derivative first perturbs the signaling network. Finally, we propose an evidence framework for distinguishing direct target modulation from secondary phosphorylation changes associated with oxidative stress, organelle dysfunction, or advanced cellular injury. Viewing OA derivatives through the kinase–phosphatase interface may provide a more precise basis for their mechanistic evaluation and future development in cancer research.

Abstract: Signal peptide peptidase-like 2A (SPPL2A) is an intramembrane protease identified as a binding partner of intraflagellar transport protein 27 (IFT27), suggesting a potential role in sperm flagellum development. However, the localization and trafficking of SPPL2A during spermiogenesis remain unknown. Here, we generated an endogenous SPPL2A–mCherry knock-in mouse model using CRISPR/Cas9 to visualize SPPL2A in living male germ cells. Expression of the fusion protein was confirmed by Western blotting, and fertility, spermatogenesis, sperm morphology, and motility were evaluated. Live-cell fluorescence imaging, immunofluorescence, and single-particle tracking (SPT) were used to characterize SPPL2A localization and trafficking. Homozygous knock-in mice exhibited normal fertility, spermatogenesis, and sperm parameters, indicating that the mCherry fusion did not disrupt SPPL2A function. SPPL2A–mCherry was detected as punctate structures in developing germ cells and elongating sperm tails. Its localization changed dynamically during spermiogenesis and was closely associated with developing tail microtubules. Live-cell SPT demonstrated bidirectional movement of SPPL2A-containing particles within developing sperm tails. Mean-squared displacement analysis indicated predominantly confined diffusion with a confinement length scale of 153 ± 15 nm, while particle trajectories showed a slight bias away from the cell body. Particle mobility progressively decreased during sperm maturation, with highly mobile puncta in early spermatids becoming largely immobile in mature sperm. These findings establish the first endogenous SPPL2A fluorescent knock-in mouse model and reveal that SPPL2A undergoes regulated intraflagellar trafficking during spermiogenesis. This model provides a valuable platform for investigating protein transport during sperm flagellum assembly.

Abstract: Induced pluripotent stem cells (iPSCs) are prone to genomic instability during prolonged culture, with recurrent chromosomal aberrations conferring selective advantages. Replication stress is a major driver of this instability, yet the repertoire of replication stress‑sensitive loci in iPSCs remains largely unexplored. Here, we mapped aphidico-lin‑sensitive fragile sites (asFS) in three independent iPSC lines using classical cytogenetic break analysis combined with Monte Carlo simulation and MiDAS mapping directly on banded metaphase chromosomes. We identified 28 asFS, which segregated into a highly active Major cluster (8 sites, accounting for 59% of breaks among asFS) and a less active Minor cluster (20 sites). Five universal asFS (9p21, 6q25-26, 20p11-12, 10q22, Xq25) were present in all three lines, representing a tissue‑specific fragility signature of pluripotent stem cells, with Xq25 shifting into the Major cluster after correction for X chromosome dosage. Minor asFS showed preferential co‑localization with physical breakpoints or minimal overlapping regions of recurrent culture‑acquired aberrations, including 20q11.21 (BCL2L1), 1q32 (MDM4), 8q24 (MYC), 17q21 (WNT3‐WNT9B), and 18q21 (DCC/FRA18B). MiDAS mapping validated most asFS and revealed additional replication stress‑sensitive loci in pericentromeric and subtelomeric regions that are difficult to score by conventional G‑banding. Comparison with fragile site maps from other cell types con-firmed tissue specificity, showing that the iPSC asFS repertoire is distinct in rank order and relative activity. Collectively, our findings indicate that the asFS repertoire in iPSCs is hierarchically organized into a stable universal core and a variable peripheral component, and suggest that Minor asFS may serve as initial substrates for adaptive culture‑acquired rearrangements. This work provides a framework for understanding how replication stress and clonal selection shape the mutational landscape of pluripotent stem cells

Abstract: Mammalian cells contain numerous membrane-bound organelles, of which endosomes serve as the initial destination for endocytosed molecules. Therapeutic agents are also internalized by cells and transported to endosomes or phagosomes, and subsequently delivered to lysosomes for degradation. Therefore, these agents require drug delivery systems (DDSs) that enable their escape from endosomes into the cytosol before lysosomal degradation; however, endosomal escape is a major limitation of current DDSs. Studies of bacterial phagosomal escape have revealed mechanisms by which host cells detect damage to organelle membranes. These membrane damage-sensing molecules also recognize membrane damage caused by artificial DDSs or physical energy-based insults. In this review, we summarize molecular mechanisms underlying the early stages of membrane damage in the plasma membrane, lysosomes and bacteria-containing vacuoles (BCVs) to better understand the early stages of endosomal membrane damage in the absence of pathogens. We summarize recent advances in galectins, endosomal sorting complexes required for transport (ESCRT) complexes, sphingomyelin, stress granules, phosphatidylinositol 4-phosphate (PI4P) at membrane contact sites, as well as annexins. We also discuss the recruitment kinetics of these molecules to damaged membranes. Although the recruitment kinetics vary depending on cell type and experimental conditions, this information provides a timeframe for the events following membrane damage, including damage sensing, membrane repair, and degradation of damaged organelles. We also discuss a potential fourth event, fusion between the plasma membrane and endosomes or lysosomes for membrane repair in annexin section. Finally, we summarize approaches for inducing “sterile” endosomal membrane damage. Future development of these approaches may facilitate the design of novel DDSs and physical energy-based strategies for manipulating specific organelles.

Abstract: Microscopy is evolving from a descriptive to a predictive tool in biology. Trained on time-lapse images, deep learning can forecast whether a cell will divide, die, or differentiate from how it looks and moves, in some systems, hours or even generations before the usual molecular markers appear, and without added fluorescent labels. Two questions frame this review and stay largely open: which image features actually carry the predictive signal, and whether a model foresees a genuinely future outcome or instead reads a state the cell has already entered. We organise the field by the visual signal that carries fate: changes in cell and nuclear shape over time, changes in brightness and texture, the dynamics of differentiation and competition, and patterns of movement. For each, we ask what must be measured, and over what area and time window, to predict fate. We then set out the main families of models, from networks that read a single image to those that read sequences of images over time and those that compress images into compact numerical summaries, outlining their strengths and limitations. Our central argument is that prediction has outrun validation: most reported performance is checked retrospectively against endpoint markers instead of on genuinely future cells. We close on the shared datasets, benchmarks, and interpretability work this gap demands, and on underexplored fates such as migration and senescence.

Abstract: Variant-based pathogenicity predictors such as REVEL evaluate missense variants in isolation, discarding the gene-length and allele-frequency context needed to compare collections of genes. In this paper, we introduce a composite gene-level metric integrating Hardy-Weinberg heterozygosity, coding-sequence length, and REVEL scores, evaluated on 55 high-confidence autism genes against the 1000 Genomes reference. It identifies elevated pathogenic burden in 48 of 55 genes, removes gene-length and variant-count confounds, and outperforms REVEL-only scoring by a large margin. Bootstrap resampling and a label-permutation control confirm the enrichment is stable and not an artefact of the scoring construction. We present it as a complementary gene-level layer for case-control and gene-set comparisons, with a nonlinear successor outlined as future work.

Abstract: Background: Piezo1 is a mechanosensitive Ca²⁺-permeable ion channel that plays a crucial role in the Ca²⁺ signaling processes of human red blood cells (RBCs). Ca²⁺ influx through Piezo1 controls the activity of various molecules that are crucial for RBC physiology and pathophysiology, including Ca²⁺-activated K⁺ channels of intermediate conductance (KCa3.1, or Gardos channels), ANO6 lipid scramblases and pannexins-1 (Panx1). Erythropoiesis, the differentiation of erythropoietic stem cells in the bone marrow to myeloid progenitor cells and then to mature (RBCs, is traditionally studied using different blood cell lines as models. Among them, K562 cells, a human chronic myeloid leukemia cell line, are multipotent progenitors of hematopoietic cells that can be differentiated along the erythroid lineage, thus making these cells an invaluable model for studying human erythropoiesis. As K562 is an immortalized cell line obtained from a specific donor, the putative differences in the relevant pathways between K562 cells and human RBCs should be identified and taken into account. Here, we aimed to reveal and compare the functional interactions between Ca²⁺ influx through Piezo1 and Gardos channel, ANO6 and Panx1 activities in K562 cells and RBCs. Methods: Database analysis was performed to confirm the expression of genes of interest and putative mutations in K562 cells. Mutations were confirmed using Sanger sequencing. Piezo1 activity in the plasma membrane was stimulated by a selective Piezo1 agonist, Yoda1. KCa3.1 activation was detected using light microscopy and single-channel patch-clamp analysis. PS exposure was detected by AnV fluorescent staining. Presence of Panx1 was shown using RT-PCR, Western Blot and immunofluorescence. K562 differentiation was induced by cytarabine-hemin treatment and confirmed by a benzidine test. Panx1 activation was probed using dye uptake assay and whole-cell patch-clamp recordings. Results: We found that, similar to RBCs, both Piezo1-KCa3.1 and Piezo1-ANO6 functional links are present in K562 cells, whereas the Piezo1-Panx1 axis is not functional because Panx1 cannot be activated by Ca²⁺ in this cell line. Conclusions: Thus, the physiologically relevant pathways associated with Ca²⁺-induced Panx1 activity in K562 cells may significantly differ from those that are functional in human RBCs.

Abstract: Caffeine exposure during pregnancy has been associated with developmental abnormalities; however, the combined effects of caffeine and glucose on embryogenesis remain unclear. In this study, zebrafish embryos were exposed to caffeine or caffeine plus glucose co-treatment from 3 hours post-fertilization to 96 hours post-fertilization to evaluate developmental, physiological, and transcriptomic changes. Morphological analyses revealed developmental abnormalities in treatment groups, including pericardial edema, abnormal body curvature, delayed tail extension, and enlarged yolk sac morphology. Significant alterations in hatch rate and heart rate were also observed following treatment exposure. RNA sequencing analysis demonstrated that caffeine induced broad transcriptomic disruption involving neuronal signaling, embryonic development, metabolism, mitochondrial activity, immune regulation, and protein turnover pathways. Compared with caffeine treatment alone, glucose co-treatment reduced the magnitude of differential gene expression and modulated developmental and metabolic signaling pathways. Alternative splicing analysis further identified widespread exon-skipping events and altered splicing patterns in genes associated with developmental and neuronal regulation. Collectively, these findings demonstrate that glucose supplementation partially reshapes caffeine-induced developmental and transcriptomic dysregulation during zebrafish embryogenesis.

Abstract: Background: The mammalian gene encoding OCT4, POU5F1, lies in or immediately adjacent to the major histocompatibility complex (MHC) region. In humans, POU5F1 is located at 6p21.33 within the extended HLA/MHC genomic neighborhood; in mice, Pou5f1 is on chromosome 17 within the homologous MHC-linked region. Marsupial data further suggest that this association belongs to an ancient mammalian immune supercomplex rather than to a lineage-specific accident. Yet birds and teleost fish show divergent arrangements, indicating that OCT4-MHC linkage is not required for all vertebrate zygotic genome activation. Objective: This paper expands a working outline on OCT4-MHC linkage into a formal theoretical article. It asks whether the tight mammalian linkage between a core pluripotency regulator and the principal immune-recognition complex could represent an evo-devo adaptation that coordinates developmental ignition, stress survival, maternal-fetal immune tolerance, chromosome-scale timing, and long-range chromatin topology. Methods: We synthesize comparative genomics, mammalian zygotic genome activation (ZGA), MHC evolution, preimplantation immunology, Hsp70 stress biology, ASAR6 replication-timing literature, 3D genome topology, and fractal genome concepts. We formulate quantitative descriptors for synteny strength, chromatin topological coupling, developmental timing, immune silencing, and evolutionary retention. Representative mammalian and non-mammalian model organisms are summarized, and a falsifiable validation pipeline is proposed. Results/Framework: The OCT4-MHC region is interpreted as a mammalian “start-and-shield” hub. OCT4/POU5F1 provides a developmental ignition module; MHC class III stress-response genes, including Hsp70 family genes, provide early cytoprotection; classical and non-classical MHC genes provide a tunable immune-recognition module; chromosome-scale elements such as ASAR6 suggest autonomous timing control; and 3D genome architecture provides a topological medium through which these modules may be co-regulated. Mathematical formulas define normalized synteny distance, linkage conservation, topological contact kernels, activation-repression coupling, fractal contact scaling, fitness effects, and Bayesian model validation. Conclusions: The OCT4-MHC linkage is best treated neither as a proven universal “origin of life” mechanism nor as a meaningless chromosomal accident. A more defensible hypothesis is that mammals conserved a genomic architecture in which the laws of development and the laws of immune recognition are compressed into a shared chromosomal neighborhood. At the most abstract level, the arrangement may reflect an isomorphic mapping between physical constraints of a genome universe and evolutionary constraints of mammalian life: stable development requires a coordinated geometry of ignition, protection, recognition, timing, and restraint.

Abstract: Background: Mowat-Wilson syndrome (MWS) is a complex neurodevelopmental and dysmorphic genetic disorder caused by heterozygous loss-of-function variants in the Zinc Finger E-Box Binding Homeobox 2 (ZEB2) gene. Methods & Objective: This comprehensive MWS study examined 301 ClinVar ZEB2 variants with 226 classified as pathogenic and 75 as likely or conflicting pathogenic including mutations, deletions, and insertions in the exonic, intronic, and splice acceptor/donor regions. The study focused on the distribution of these MWS variant sets across the ZEB2 gene, especially within its six functional domains. We used the STRING protein-protein interaction network to infer the molecular and biological functions of the ZEB2 protein. Results & Conclusions: The findings challenge the assumption that incidence of MWS pathogenicity is lower with variants located toward the C-terminal end with intact functional ZEB2 domains and revealed truncating variants near the N-terminus as not significantly different from those near C-terminus. Pathogenicity within functional domains is notably higher. The highest per-amino-acid pathogenicity was found with C-ZF type 6 Zfn, further indicate that maintaining integrity of C-terminal cluster of three ZFNs is equally crucial for this protein to bind bipartite CACCT elements. Noticeable increases are seen in pathogenicity within the SMAD-MH2-binding domain, DNA-binding/homeodomain, CtBP-interacting domain, and C-ZF Zfn types 6-7. Overall, pathogenicity in domain regions was 4.1% higher than in non-domain regions. No significant differences were observed in occurrence of frameshift, nonsense, and missense variants among these two groups of cases. More than 4% of pathogenic cases and over 10% of likely/conflicting pathogenic cases are caused by variants in intronic, splice-acceptor, or splice-donor sites, requiring further studies including clinical investigations Analysis of STRING interactions for ZEB2 identified key molecular and biological functions likely to be affected in this complex neurodevelopmental and dysmorphic genetic disorder and useful for clinical assessment.

Abstract: Hybridoma technology remains one of the most reliable and widely used platforms for generating highly specific monoclonal antibodies for use in diagnostics, fundamental research, and clinical practice. Moreover, the combination of unlimited proliferative capacity with the preservation of a key B lymphocyte function, antibody production, enables hybridoma cells to support a comprehensive functional evaluation of cellular responses. Thus, by integrating analysis of proliferation, viability, and productivity, hybridoma-based approaches enable the detection of differential modulatory effects and offer a nuanced assessment of compound bioactivity. This review provides a comprehensive analysis of the functional parameters of hybridoma cells: viability, proliferation, and productivity, as well as the methods used for their evaluation. The main stages of hybridoma cell generation, advances in hybridoma technology, and current applications of hybridoma cells are also reviewed. A key aspect of this review is the differential modulation of functional parameters of hybridoma cells. Modulation of culture conditions and bioactive compounds can differentially influence growth dynamics and specific antibody yield, often revealing an inverse relationship between proliferation and productivity.

Abstract: Background: Population aging is driving a convergent rise in three disorders historically managed in isolation: cardiovascular disease, neurocognitive decline, and osteoporotic bone loss. Accumulating mechanistic data indicate that these systems are coupled through shared regulators of calcium trafficking, inflammation resolution, vascular integrity, and inflammaging, supporting the concept of an integrated "Heart-Brain-Bone" axis. Three lipid-soluble nutrients-long-chain omega-3 polyunsaturated fatty acids (EPA/DHA), vitamin D3 (cholecalciferol), and vitamin K2 (menaquinone-7 [MK-7])-act on overlapping nodes of this network. Methods: We conducted a structured narrative review of mechanistic, observational, and randomized controlled trial (RCT) evidence, grading the strength of support for each claim. Results: Vitamin K2-dependent gamma-carboxylation of matrix Gla protein (MGP) and osteocalcin is proposed to reciprocally direct calcium away from the arterial wall and toward the bone matrix, offering a candidate mechanistic resolution of the "calcium paradox" associated with isolated vitamin D3 supplementation; EPA/DHA-derived specialized pro-resolving mediators may support resolution of endothelial and neuronal inflammation; and bone-, vascular-, and brain-derived signals (osteocalcin, FGF23, the neurovascular unit) interconnect the three organs. Human clinical evidence, however, is heterogeneous and formulation- and population-dependent: cardiovascular omega-3 RCTs are discordant (REDUCE-IT, which used icosapent ethyl [an EPA ethyl ester], positive; VITAL/STRENGTH/ASCEND null, predominantly in lower-risk or replete cohorts), cognitive trials are largely null or subgroup-dependent (MAPT, DO-HEALTH), and MK-7 improves surrogate bone and calcification biomarkers, with only a single recent hard-endpoint coronary calcification RCT (VitaK-CAC). Conclusions: The combined use of long-chain omega-3, vitamin D3, and MK-7 is mechanistically rational and hypothesis-generating rather than clinically established; benefit appears most plausible in individuals with elevated risk or demonstrable nutritional insufficiency, and least in replete, low-risk populations. Findings should be interpreted within a broader healthy-aging context that includes lifestyle and psychosocial factors. Adequately powered factorial RCTs stratified by baseline Omega-3 Index, 25(OH)D, and vitamin K status, with prespecified mechanistic biomarkers and hard endpoints, are required.

Abstract: Cardiolipin (CL), a unique dimeric phospholipid with four acyl chains and a characteristically small polar head group, stands as one of the most compelling examples of evolutionary continuity in cell biology. Present in the plasma membrane of a-proteobacteria and conserved without fundamental modification in the inner mitochondrial membrane (IMM) of all eukaryotes examined, CL has been retained across approximately two billion years of evolution, a period over which the mitochondrion shed thousands of its original genes. This degree of conservation demands an explanation that transcends structural necessity alone. Here we propose, and document with biochemical and cell biological evidence, that CL functions as a programmable signaling hub: a lipid species whose physical chemistry and membrane address allow it to assemble distinct supramolecular platforms in response to discrete stress signals, each platform transducing a specific mitochondrial state into a defined cell fate outcome. Three core CL signaling platforms are described. Platform 1, the catalytic peroxidase platform, converts the constitutive CL–cytochrome c (cyt c) structural complex into an enzymatic reaction under oxidative stress, generating oxidized CL (oxCL) species that commit the cell to apoptosis by releasing cyt c from the IMM. Platform 2, the receptor-like mitophagy platform, exploits NME4-dependent CL scramblase activity to translocate CL from the IMM to the outer mitochondrial membrane (OMM) surface upon membrane potential dissipation, creating an externalized "eat-me" signal that LC3-II on autophagic membranes recognizes directly. Platform 3, the caspase-8/BID activation platform, assembles a CL microdomain scaffold at the OMM that recruits caspase-8 from death receptor complexes, accelerates Bid cleavage by three orders of magnitude, and couple extrinsic apoptotic signals to mitochondrial outer membrane permeabilization (MOMP). An emerging fourth axis links CL externalization to innate immune activation via NLRP3 inflammasome recruitment. We argue that the deep evolutionary conservation of CL reflects not its structural roles per se, but the irreplaceable nature of these signaling functions. Functions already present in ancestral bacteria and progressively elaborated as eukaryotic cell death, quality control, and immunity coevolved with the organelle itself.

Abstract: We report here on an attempt to create an engineered structure which can reproduce the physical and mechanical properties of the land plant sporangium. This work is part of a broader effort to understand the stress mechanical environment of the land plant sporangium and its role in initiating reproductive development. A second purpose is to extend the range of experimental methods and tools available for the study of the physical environment of plant cell growth, and the mechanics of trans-cellular signaling in plant development. We describe an experimental protocol, based on the microfluidic encapsulation of living plant protoplasts in multilayered microbeads composed of tunable hydrogel materials whose mechanical properties can be modified to mimic the stress-mechanics of the living sporangium. Our results demonstrate the successful encapsulation of living plant protoplasts in dual-layered microbeads consisting of an inner layer of gelled agarose and an outer layer of cross-linked alginate/methacrylate.

Abstract: In addition to soluble biochemical factors, biophysical cues from the extracellular matrix (ECM) can play an integral role in regulating cell behavior. Previous work has demonstrated that varying ECM dimensionality (3D vs. 2D) and structure (fibrillar vs. non-fibrillar) can produce distinct cell phenotypes; however, the transcriptional changes underlying how cells sense and respond to these different environments are not well understood. Here, we used bulk RNA sequencing to compare the gene expression profiles of primary rabbit corneal keratocytes cultured on collagen-coated substrates and on top of or embedded within fibrillar collagen matrices under serum-free conditions. Differential expression and functional analyses revealed distinct transcriptional profiles across the culture conditions and identified differentially expressed genes and enriched signaling pathways primarily related to the ECM, cell-matrix interactions, cell mechanics, and proliferation. Cells cultured in 3D fibrillar conditions exhibited gene expression patterns that better reflected a quiescent in vivo keratocyte phenotype, including broad suppression of proliferation and ECM synthesis-related genes. In contrast, cells cultured on 2D substrates showed higher expression of genes associated with an activated phenotype. Overall, these findings demonstrate the strong influence of biophysical cues from the ECM on keratocyte gene expression and highlight the importance of selecting physiologically relevant in vitro models for studies of corneal cell biology, wound healing, and regenerative therapies.

Abstract: Glioblastoma remains the most lethal primary malignancy of the central nervous system, and the modest gains achieved with maximal surgery, radiotherapy and temozolomide have not been matched by the immune checkpoint inhibitors and antigen-specific vaccines that reshaped the treatment of many extracranial cancers. The recurrent disappointment of these approaches has been attributed less to a single molecular lesion than to a confluence of obstacles: profound intratumoural heterogeneity, a densely immunosuppressive and myeloid-rich microenvironment, sequestration and exhaustion of conventional T cells, and the practical difficulty of delivering effectors across the blood-brain barrier. Against this background, γδ T cells have attracted interest as an unconventional effector population that recognises transformed cells through stress-associated and metabolic cues rather than peptide-major histocompatibility complex (MHC) complexes, that kills in an MHC-unrestricted manner, and that can be expanded from healthy donors for allogeneic, off-the-shelf use with little expectation of graft-versus-host disease. This narrative review examines, with a deliberately critical lens, the biological rationale and the experimental evidence for γδ T-cell-based immunotherapy of glioblastoma. We summarise the developmental biology and functional subsets of human γδ T cells, the NKG2D-, DNAM-1- and T-cell-receptor-dependent mechanisms through which they engage glioma cells and glioma stem-like cells, and the in vitro and animal-model studies that underpin the field, taking care not to overstate efficacy that has so far been demonstrated only in preclinical or early-phase settings. We then weigh the principal opportunities—locoregional and repeated dosing, combination with chemoradiotherapy, checkpoint blockade and antibody-based redirection—against barriers that include limited persistence, uncertain intratumoural trafficking, donor and manufacturing variability, and the unsettled requirements of potency testing and trial design. Throughout, γδ T cells are presented as a biologically plausible but still investigational strategy whose clinical value will be determined by adequately powered trials rather than by mechanistic appeal alone.

Abstract: Background: Keratoconus is a progressive corneal ectasia characterised by extracellular matrix (ECM) loss and an emerging inflammatory component, for which no disease-modifying molecular therapy exists. Exosomes derived from limbal and mesenchymal stem cells are an attractive cell-free therapeutic modality, but the cargo that should be delivered is undefined, and no curated limbal stem-cell (LSC) exosome cargo dataset currently exists. Methods: We reanalysed a public keratoconus corneal RNA-sequencing dataset (GEO: GSE77938; discovery and replication cohorts) with DESeq2, defined a replicated differentially expressed gene (DEG) set, and performed Gene Ontology, KEGG and Reactome enrichment. A high-confidence protein-protein interaction (PPI) network (STRING) identified hub genes. We integrated keratoconus disease-gene evidence (Open Targets Platform) and documented extracellular-vesicle cargo (ExoCarta, Vesiclepedia) and computed a transparent Cargo Prioritization Score (CPS) to nominate candidate LSC-exosome therapeutic cargo. Results: 1677 DEGs were detected in discovery (152 up, 1525 down) and 1380 were replicated. Enrichment was dominated by extracellular matrix organization; adaptive immune response; mononuclear cell differentiation. Network analysis nominated ECM and immune hub genes. The CPS prioritised COL1A1, FN1, COL4A1, COL3A1, COL5A1, MMP1 as leading restoration-cargo candidates, all documented as EV cargo and present in the mesenchymal stem-cell EV reference proteome. Conclusions: This fully reproducible, real-data framework provides a ranked, evidence-traceable shortlist of candidate LSC-exosome cargo for keratoconus and an explicit account of current data gaps to guide experimental validation.

Abstract: Ribonucleoprotein granules such as processing bodies (P-bodies) and stress granules (SGs) are membrane-less organelles that regulate mRNA metabolism through liquid–liquid phase separation. UBAP2L drives SG assembly and can bridge P-bodies with SGs, yet how it is mobilized between these compartments remains unclear. Here, using co-immunoprecipitation, GST pull-down, CRISPR-Cas9-mediated knockout, and immunofluorescence microscopy, we demonstrate that CCHCR1 directly binds UBAP2L and that this interaction is dynamically regulated by stress intensity. Under mild oxidative stress, CCHCR1 retains UBAP2L in P-bodies; as stress intensifies, this interaction weakens, permitting UBAP2L release for SG assembly. CCHCR1 deficiency aberrantly traps UBAP2L in P-bodies via enhanced DDX6 association, resulting in defective SG assembly, delayed maturation, and increased P-body–SG fusion. These findings establish CCHCR1 as a stress-responsive switch that controls UBAP2L partitioning between P-bodies and stress granules, thereby controlling the threshold and kinetics of SG biogenesis.

Abstract: The developmental basis of retroperitoneal fascial lamination remains unresolved, as classical peritoneal fusion theories and fat-compaction models cannot fully explain the consistent formation of the anterior and posterior renal fasciae or their behavior in cases of congenital renal agenesis. To clarify the underlying mechanobiology, we conducted a retrospective radiological analysis of unenhanced computed tomography (CT) scans, including rare cases of unilateral renal agenesis, interpreting fascial configurations within a framework incorporating tension-driven lamination, orthogonal Poisson compression, and subtraction-based reasoning. Across all cases, a continuous fascial plane was unequivocally preserved at the predicted anatomical location of the parietal lamina of the posterior renal fascia despite the lifelong absence of the kidney. However, the renal-vacant side consistently exhibited reduced total fascial thickness (mean 1.52 mm vs. 1.85 mm). This asymmetric thinning aligns with the selective absence of the organ-dependent inner lamina and the preservation of a system-derived parietal lamina established during mid-gestation. From a comparative developmental perspective, whereas quadrupedal precocial mammals preferentially allocate fetal movement energy to appendicular stiffening while maintaining a compliant trunk (heterochrony), humans front-load truncal stiffening (an evolutionary shift toward earlier-than-expected ossification timing) to protect a geometrically vulnerable pelvic architecture associated with obligate bipedalism. The convergence of this human-specific "early-completed truncal box" and powerful fetal movement energy concentrates orthogonal Poisson compression (a biological "ironing" effect) in the retroperitoneum, triggering synchronous multilaminar sheet formation. These findings support an evolutionary and mechanobiological model in which the laminated architecture of the retroperitoneal fascia emerges from the interplay of geometric scaling, the human-specific timing of skeletal stiffening, and multiaxial tension fields.