Search Results (894)

Background: Microtubules play a crucial role in neuronal structure and function, with their stability and dynamics regulated by posttranslational modifications (PTMs) such as polyglutamylation. In Alzheimer’s disease (AD), the microtubule-associated protein TAU becomes mislocalized into the somatodendritic compartment (‘TAU missorting’), dissociates from microtubules, aggregates into neurofibrillary tangles, and contributes to microtubule destabilization and neuronal death. Objectives and Methods: Here, we investigated the role of tubulin tyrosine ligase-like proteins (TTLLs) in TAU missorting and microtubule dysregulation using human-induced pluripotent stem cell (hiPSC)-derived cortical neurons treated with oligomeric amyloid-beta (oAβ) to replicate AD-like conditions. TTLL1, TTLL4, and TTLL6 were selectively knocked down (KD) to assess their impact on TAU missorting and microtubule stability. Fluorescence resonance energy transfer (FRET) microscopy was used to examine proximities between TAU and TTLL proteins. Results: We observed TAU missorting, increased tubulin polyglutamylation, decreased tubulin acetylation associated with microtubule destabilization, and synaptic declustering in oAβ-treated neurons. TTLL1 KD significantly reduced TAU missorting, tubulin polyglutamylation, and synaptic disintegration, while TTLL4 KD showed moderate effects, and TTLL6 KD restored microtubule acetylation. Importantly, TTLL KD did not impair neuritic networks, dendritic complexity, or neuronal activity. FRET microscopy in HEK293T cells revealed a close molecular proximity between TAU and TTLL1 consistent with a potential direct or complex-mediated association, but not with other TTLLs, suggesting a direct role of TTLL1 in TAU-mediated toxicity. Conclusions: Our findings identify TTLL1 as a promising therapeutic target for limiting TAU-associated cytoskeletal pathology in AD. These results support further development of pharmacological or genetic strategies targeting TTLL1 as a disease-modifying approach for AD and related tauopathies. Full article

Ischemic heart disease is currently diagnosed mainly through cardiac computed tomography (CT) angiography and functional testing, both of which detect only advanced arterial macrocalcification, at a stage when treatment can merely slow disease progression rather than reverse it. Yet, macrocalcification represents the end product of a much earlier molecular process, which is microcalcification. This process is driven by smooth muscle cell and macrophage apoptosis, matrix vesicle release, and osteogenic phenotypic transitions within the arterial intima, occurring years to decades before mineral deposits become visible on CT. [¹⁸F]Sodium fluoride (NaF) positron emission tomography (PET) exploits fluoride binding at accessible hydroxyapatite surfaces to detect increased tracer uptake associated with active mineral deposition, including mineralization occurring at a microscopic scale below the direct spatial resolution of clinical PET. Studies demonstrate that anti-atherosclerotic interventions, including statins, and tissue-nonspecific alkaline phosphatase inhibition can suppress NaF uptake even when CT-based calcium scores remain unchanged or continue to rise, a dissociation now also observed in human trials of statins and PCSK9 inhibitors. This review traces the cellular and histological basis of arterial calcification, outlines the principles and limitations of NaF-PET imaging, and evaluates its emerging role—supported by artificial intelligence-based quantification—as a tool for monitoring targeted anti-atherosclerotic treatment. Full article

Background: Therapeutic vaccination against human papillomavirus type 16 (HPV16) is frequently limited by the weak HLA-A*02:01 presentation of native E6/E7 oncoprotein epitopes. This study aims to utilize an integrated computational-experimental pipeline to design and evaluate anchor-optimized altered peptide ligands (APLs) for improving peptide presentation and HPV16-specific T-cell responses. Methods: Anchor-residue substitutions were introduced into three wild-type E6/E7 epitopes. Candidates were prioritized in silico based on predicted presentation, affinity, immunogenicity, and toxicity, and subsequently evaluated via in vitro functional assays using HLA-A*02:01-positive donor cells and molecular dynamics (MD) simulations. Results: Anchor optimization successfully converted weak binders into strong binders; for instance, E6apl improved predicted affinity from 329.33 to 6.21 nM. Crucially, candidate E7apl1 exhibited normal CD8⁺ T-cell expansion but reduced IFN-γ secretion, revealing a distinct binding–immunogenicity dissociation. MD simulations suggested that altered peptide conformational dynamics may contribute to differences in functional activity. Subsequently, an optimized six-peptide formulation (three APLs and three wild-type epitopes) was assembled. The resulting multi-epitope HPV-specific cytotoxic T lymphocytes (meHPV-CTLs) mediated target-specific cytotoxicity against cervical cancer cells, achieving 74.1% ± 5.1% specific lysis at an effector-to-target ratio of 30:1, which was largely abrogated by HLA class I blockade. Conclusions: These proof-of-concept findings demonstrate that stable peptide–MHC binding is a necessary but insufficient condition for optimal T-cell activation. The experimentally characterized multi-epitope formulation provides a proof-of-concept strategy for further preclinical evaluation of HPV16-targeted peptide-based immunotherapies. Moreover, because these anchor-optimized APLs are defined at the sequence level, these sequence-defined APLs may potentially be explored in alternative vaccine delivery platforms, including mRNA-based approaches, in future studies. Full article

Preclinical models play a critical role in the development of PSMA-targeted radiopharmaceuticals for prostate cancer. However, many existing models have practical limitations, including slow tumor growth, low engraftment rates, and restricted availability, and all human PSMA-positive lines are confined to immunodeficient hosts. We developed and characterized a novel PSMA-expressing transgenic cell line, CT26-PSMA, as a practical tool for preclinical screening of PSMA-targeting agents. The CT26-PSMA cell line was established by stable transfection of the murine colon carcinoma CT26 cell line with human PSMA using the Sleeping Beauty transposon system. PSMA expression was confirmed by RT-qPCR (reverse transcription quantitative polymerase chain reaction), flow cytometry, and radioligand saturation binding on intact cells. Two [⁶⁸Ga]Ga-labeled radioconjugates—the well-established PSMA-617 and a newly synthesized conjugate (Conjugate-1)—were used to validate the functionality of the model through in vitro binding, uptake and internalization studies, and through ex vivo biodistribution in CT26-PSMA tumor-bearing athymic male nu/nu mice. The CT26-PSMA cell line demonstrated high and stable PSMA expression, with approximately 95% of cells expressing the biomarker and no measurable loss over 16 passages in antibiotic-free medium. Saturation binding gave a receptor density of ∼3.5 × 10⁶ sites per cell, approximately four-fold higher than that of LNCaP cells (∼0.8 × 10⁶), with dissociation constants that were indistinguishable between the two radioconjugates and between the two cell lines (K_d 9.0–11.6 nM). Subcutaneous tumors reached ~300 mm³ within 8–10 days of inoculation, with a take rate of 10/10 versus 1/10 for LNCaP (Fisher’s exact test, p = 1.2 × 10⁻⁴). Both radiotracers showed saturable, 2-PMPA-blockable binding and uptake in CT26-PSMA cells, confirming the functional activity of the recombinant receptor. Biodistribution studies revealed accumulation of both conjugates in CT26-PSMA tumors, with generally comparable tumor-to-background profiles. The CT26-PSMA cell line represents a robust, rapid, and reproducible platform for preclinical evaluation of PSMA-targeting radiopharmaceuticals, and its murine BALB/c origin permits engraftment in immunocompetent or minimally immunosuppressed hosts, whereas existing human PSMA-positive lines do not. It is intended as a screening platform rather than as a model of prostate cancer biology. The validation data obtained with [⁶⁸Ga]Ga-labelled conjugates confirm the suitability of this cell line for future studies of PSMA-directed compounds. Full article

B cell aplasia (BCA), the sustained depletion of circulating CD19-positive B cells, is the defining on-target, off-tumor consequence of anti-CD19 chimeric antigen receptor (CAR) T cell therapy. Despite its occurrence across all approved CAR T cell products and all responding patients, BCA has not been systematically reviewed as a standalone clinical and biological phenomenon. This review synthesizes data from landmark trials and real-world cohorts across B cell-acute lymphoblastic leukemia (B-ALL), large B cell lymphoma, follicular lymphoma, mantle cell lymphoma, and chronic lymphocytic leukemia to characterize BCA incidence, recovery kinetics, prognostic significance, and management implications. Anti-BCMA products, which mechanism of action differs fundamentally, depleting plasma cells rather than B cell precursors, are intentionally excluded from this review. BCA is observed in all responders and is consistently absent in non-responders across reported cohorts , making it a reliable pharmacodynamic marker of CAR T cell activity. Its prognostic significance is disease-specific: in lymphoma, BCA recovery does not predict relapse and durable remission is achievable independent of sustained aplasia; in B-ALL, early BCA recovery within six months is a robust independent predictor of CD19-positive relapse, while persistent BCA correlates with sustained remission. CD19-negative antigen-escape relapse occurs preferentially in the presence of intact BCA and high pre-infusion tumor burden. Combining BCA kinetics with bone marrow next-generation sequencing minimal residual disease assessment at day 28 and month 3 constitutes the most powerful post-infusion risk-stratification framework currently available. BCA is mechanistically dissociated from hypogammaglobulinemia: IgM declines rapidly and profoundly, IgA more slowly, while IgG—maintained by CD19-negative long-lived plasma cells—is the most preserved isotype. No B cell count threshold below which hypogammaglobulinemia becomes clinically significant has been established. Evidence-informed IVIG replacement thresholds are proposed, though no randomized trial data exist to support them, representing a critical gap requiring prospective investigation. Full article

The practical viability of lithium–sulfur batteries (LSBs) is severely hindered by sluggish liquid–solid conversion kinetics and the polysulfide shuttle effect. Herein, we report an in situ-derived ferroelectric-semiconductor Bi₄Ti₃O₁₂-Bi₂S₃ heterojunction as a multifunctional separator coating. The intimate atomic-level coupling at the heterointerface generates a built-in electric field that, synergizing with the spontaneous ferroelectric polarization of Bi₄Ti₃O₁₂, structurally intensifies polysulfide chemisorption and lowers the activation energy for bi-directional Li₂S precipitation/dissociation. Furthermore, the localized polar field appears to homogenize lithium-ion flux, which may contribute to improved lithium anode stability. Consequently, cells featuring the modified separator deliver a high initial capacity of 1172 mAh g⁻¹ at 0.5 C and demonstrate good cycling stability over 500 cycles with a low capacity decay rate of 0.096% per cycle. This in situ interfacial engineering offers a promising kinetic regulatory strategy for improving the performance of LSBs. Full article

Background/Objectives: Autoimmune nodopathies are a distinct subgroup of immune-mediated peripheral neuropathies caused by antibodies targeting nodal and paranodal proteins, including contactin-1 (CNTN1). These disorders are frequently misclassified as chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), despite fundamental differences in pathophysiology, clinical course, and treatment response. Growing evidence indicates that systemic manifestations, such as proteinuria, may represent relevant diagnostic red flags. We report a case of anti-CNTN1 autoimmune nodopathy with renal involvement and long-term follow-up after rituximab therapy. Methods: We describe the longitudinal clinical, electrophysiological, laboratory, and therapeutic features of a patient presenting with an acute/subacute sensory ataxic neuropathy evolving into a chronic immune-mediated disorder. Serial nerve conduction studies were performed during multiple hospital admissions and follow-up visits. Autoantibody testing for nodal/paranodal antigens was undertaken, and systemic biomarkers were monitored over time. Results: The patient initially exhibited a robust response to intravenous immunoglobulin (IVIg), consistent with an acute inflammatory neuropathy. Subsequent relapse was characterized by cranial nerve involvement, worsening sensory ataxia, peripheral edema, albuminocytologic dissociation on cerebrospinal fluid analysis, proteinuria, and an inverted albumin/gamma globulin ratio, redirecting the diagnostic hypothesis toward CIDP. The detection of anti-CNTN1 antibodies ultimately established the diagnosis of autoimmune nodopathy. Owing to poor durability of IVIg, rituximab was initiated, resulting in sustained clinical improvement and near-complete recovery of motor and sensory nerve conduction parameters at one-year follow-up (November 2025). Conclusions: This case emphasizes the diagnostic relevance of extraneurological biomarkers, including proteinuria and peripheral edema, in autoimmune nodopathies and supports early nodal/paranodal antibody testing in atypical demyelinating neuropathies. Prompt B-cell-directed therapy may enable functional recovery of nodal integrity and improve long-term outcomes. Further studies are needed to clarify potential immunological triggers, including anti-IL-23 therapies, in the pathogenesis of anti-CNTN1 autoimmune nodopathy. Full article

Scaffold architecture shapes tissue regeneration through the mechanical, topographical, and biochemical cues it presents to cells, yet geometrically elaborate scaffolds do not reliably produce more organized tissues, while comparatively simple architectures can exert strong organizational effects. We argue that scaffold performance is better understood by distinguishing geometric complexity from effective informational dimensionality: a relational property of the scaffold–cell system, defined as the number of independently manipulated architectural directions that produce distinguishable, above-noise changes in a jointly measured mechanotransductive response. Unlike structural entropy, fractal dimension, or feature-counting metrics, this construct depends on cellular accessibility, cue persistence, and non-redundancy. Mechanotransduction supplies its biological basis, integrating scaffold-derived cues through focal adhesions, cytoskeletal organization, nuclear deformation, and YAP/TAZ signaling, and we distinguish early resolvability from later organizational stabilization. We outline an operational strategy for estimating both from factorial scaffold libraries, common readout panels, and rank-based analysis of the response mapping, illustrated with selected experimental precedents rather than a systematic evidence sample. Positioned relative to biomimetic, mechanobiology-guided, and morphospace approaches, it yields testable predictions on dimensional compression, redundancy, and the resolvability–stability dissociation. Scaffold design is thus reframed from maximizing complexity or native resemblance toward engineering stable, cell-readable dimensions of organization. Full article

Bulk heterojunction (BHJ) active layers in organic photovoltaics (OPVs) are nanostructured composites in which electron-donating and electron-accepting semiconductors form interpenetrating phases for exciton dissociation and charge transport. The power conversion efficiency (PCE) of these organic-organic nanocomposites is governed by structural features spanning multiple length scales: molecular packing and energy-level alignment at donor/acceptor (D/A) interfaces, phase-separation morphology and crystallite connectivity, and thin-film optical and charge-transport characteristics. Rational design of high-performance OPV nanocomposites requires multiscale numerical modelling that bridges quantum chemistry, mesoscale morphology simulation, and device-scale optoelectronic modelling. This review surveys and critically compares recent advances in the structural design and numerical simulation of OPV BHJ nanocomposites. At the molecular scale, we examine density functional theory and non-adiabatic molecular dynamics approaches for resolving charge-separation driving forces, interfacial energy-level alignment, and exciton dynamics. At the mesoscale, we discuss molecular dynamics, kinetic Monte Carlo, and electronic coarse-graining methods for describing phase separation, crystallization kinetics, morphology evolution, and charge transport. At the device scale, we review exciton-diffusion, optical transfer-matrix, and drift-diffusion models that quantitatively link morphology to photovoltaic performance metrics. The review also evaluates how machine learning, high-throughput screening, surrogate models, and generative design accelerate donor–acceptor selection and morphology optimization, while distinguishing benchmark predictions from experimentally validated design rules. Across these scales, we compare the strengths, assumptions, and validation limits of the principal modelling approaches. Finally, we highlight emerging multiscale integration frameworks, including sequential parameter-passing pipelines and differentiable digital-twin concepts. By framing OPV BHJ layers as nanocomposites whose performance bottlenecks map onto composite-design challenges such as interface integrity, phase connectivity, multiscale charge transfer, and degradation-aware design, this review connects OPV modelling with broader structural-composites thinking for next-generation organic solar cells. Full article

Mycobacterium bovis Bacille Calmette–Guérin (BCG), the only licensed vaccine against tuberculosis, provides inconsistent protection against pulmonary tuberculosis, reflecting an incomplete understanding of how vaccine-induced immunity is organized within tissues. Emerging evidence indicates that the route of vaccination is not merely a technical variable but a critical determinant of immune programming. Whereas parenteral BCG primarily elicits systemic immune responses, mucosal delivery reprograms immunity at the respiratory interface by promoting localized trained innate immunity, tissue-resident memory T (T_RM) cells, and early containment of infection. In this review, we propose an integrated framework of “immune layering,” in which protection emerges through the coordinated interactions of epithelial regulation, trained innate immunity, tissue-resident adaptive memory, regulatory homeostasis, and systemic immune support across spatial and temporal scales. Within this framework, trained innate immunity serves as an initial conditioning layer that shapes subsequent adaptive differentiation, whereas epithelial- and microbiota-associated regulatory networks establish the tissue context in which immune responses are initiated, organized, and maintained. Importantly, effective mucosal immunity depends on a dynamically regulated equilibrium rather than maximal immune activation. The dissociation between enhanced early pulmonary immune responses and limited long-term protection underscores the influence of tissue-specific regulatory constraints and environmental context on vaccine efficacy. This framework redefines correlates of protection by identifying the vaccination route and tissue-level immune organization as fundamental determinants of protective immunity, thereby providing a conceptual foundation for the rational development of next-generation mucosal tuberculosis vaccines. Full article

Natural plant extracts rich in polyphenols are increasingly being studied as multifunctional food ingredients with antioxidant and stabilizing properties. In this study, Ocimum basilicum L. extracts were evaluated as biological cryoprotective agents for Streptococcus thermophilus. The extracts contained high levels of phenolic compounds (~1350–2200 mg GAE equivalents/L) and exhibited strong antioxidant activity (up to 5.6 mM Trolox equivalents). Density functional theory calculations showed low O–H bond dissociation energies (~72–74 kcal/mol in ethanol) for key components, including luteolin and rosmarinic acid. These calculations indicate a high hydrogen donation capacity comparable to or exceeding that of ascorbic acid. Molecular dynamics simulations demonstrated the preferential localization of major phenolic compounds at the membrane–water interface in a POPC bilayer membrane. The interaction of molecules with POPC increased membrane thickness and formed stable hydrogen-bond networks with lipid head groups. Experiments showed that systems based on basil extract significantly increased the survival of bacteria after storage at −25 °C, with the number of viable cells reaching (1.5–2.75) × 10⁸ CFU/mL. This effect was observed in comparison with control groups that used saline or sucrose. Fluorescent analysis of live/dead cells confirmed the improvement in cell membrane preservation. At the same time, no signs of metabolic inhibition were detected. Taken together, the experimental and computational results support the hypothesis that the cryoprotective effect may involve complementary antioxidant and membrane-associated interactions. However, direct biophysical validation of the proposed membrane mechanism is still required. These results emphasize that polyphenol extracts are promising natural functional ingredients for improving the stability and shelf life of probiotic and starter cultures in food systems. Full article

Poly(ethylene oxide) (PEO)-based solid polymer electrolytes are regarded as highly promising solid electrolyte materials owing to their favorable chain flexibility. However, their practical application is hindered by low room-temperature ionic conductivity and poor mechanical properties. To address these issues, a metal–organic framework (Cu-MOF) with a 2D layered structure and 1D microchannels is introduced into PEO to form a composite solid electrolyte. The results reveal that Cu-MOF can suppress PEO crystallization through steric hindrance and coordination interactions, thereby increasing the fraction of the amorphous phase. Moreover, its unsaturated metal sites can attract TFSI⁻ anions, promoting the dissociation of the sodium salt and enhancing sodium-ion transport. Theoretical calculations and molecular simulations further confirm the regulatory role of Cu-MOF in ion transport. Leveraging this mechanism, the Na₃V₂(PO₄)₃/C|PCM-8%|Na cell delivers exceptional electrochemical performance over a wide temperature range. At room temperature, the capacity exhibits virtually no decay after 200 cycles at 0.5 C, and outstanding rate capability is maintained even at a high rate of 4 C. At a temperature of 65 °C, a capacity retention of 91.4% is achieved after 200 cycles at 0.5 C. This study offers a highly promising strategy for the development of wide-temperature-range, high-performance solid-state sodium batteries. Full article

Two tri-carbonyl complexes, [Mn(CO)₃(5cpa)Br] (1) and [Re(CO)₃(5cqn)(OCH₃)] (2), were synthesized, where 5cpa is 5-Cl-2-picolinic acid and 5cqn is 5-Cl-8-Hydroxyquinoline. Their structures were determined using X-ray diffraction techniques. The electronic absorption and IR spectra of the complexes were experimentally measured and theoretically assigned through density functional theory (DFT) calculations. The photo-induced CO release was verified using time-resolved infrared spectroscopy, and the transfer of CO to hemoglobin (Hb) was monitored by UV-vis spectroscopy. The rate of CO release and transfer from Mn complex 1 is significantly faster than that from Re complex 2. Complex 2 exhibits higher cytotoxicity against HeLa cells than complex 1, with IC₅₀ values of 40.1 μM and 10.6 μM for 1 and 2, respectively, which decrease to 16.2 μM and 4.9 μM after photo irradiation. Moreover, 1 exhibited a stronger binding constant (K_b) with human serum albumin (HSA) than 2, with values of 1.6 × 10⁶ and 7.0 × 10⁵ M⁻¹, respectively. The structures of HSA complex adducts revealed that both the resulting [Mn(CO)₃(5cpa)]⁻ and [Re(CO)₃(5cqn)]⁻ group coordinate with the N atom of His146, while four additional dissociated Mn-CO groups were observed to bind to HSA for complex 1. This study provides insights into the stability, possible metabolic pathways, and potential applications of these carbonyl complexes. Full article

Sjögren’s disease (SjD) is a chronic autoimmune disease characterized by lymphocytic infiltration and dysfunction of the exocrine glands, with manifestations extending beyond glandular sicca symptoms to multiple extraglandular systems. Although the pathogenesis of SjD remains incompletely understood, growing evidence indicates that a complex cytokine network involving both innate and adaptive immune pathways plays a central role in disease development. This narrative review summarizes recent updates on cytokine signaling in SjD across three clinically relevant domains. In glandular inflammation, activation of salivary gland epithelial cells through Toll-like receptor pathways triggers type I interferon (IFN) signaling via plasmacytoid dendritic cells, while IFN-γ, Th17-related cytokines (IL-6, IL-17, IL-22), BAFF/APRIL, and chemokines (CXCL10, CXCL12, CXCL13) collectively sustain local inflammation and ectopic lymphoid organization. The BAFF/APRIL axis, a systemic type I IFN signature, and IL-21–follicular helper T cell–B cell interactions primarily drive systemic immune activation, which together underlie autoantibody production, hypergammaglobulinemia, and a lymphoma-prone phenotype. In contrast, constitutional symptoms such as fatigue, pain, and dryness frequently dissociate from classical inflammatory activity and are better explained by neuroimmune–metabolic mechanisms, including the IFN-γ–IDO–kynurenine pathway and symptom-associated proteomic signatures. Collectively, these findings underscore the heterogeneous nature of SjD, in which glandular inflammation, systemic immune activation, and constitutional symptoms are driven by distinct yet partially overlapping cytokine pathways. Recognizing this heterogeneity has direct implications for cytokine-targeted therapy, suggesting that future trials should stratify patients by disease phenotype (IFN-high, B cell-dominant, and symptom-dominant) rather than treating SjD as a uniform population. Full article

Cardiac fibroblasts (cFbs) are important in vitro models for studying myocardial fibrosis, extracellular matrix homeostasis, and tissue remodeling. However, establishing primary cultures from adult mouse hearts remains technically demanding due to the limited proliferative capacity of freshly isolated cells, their susceptibility to enzymatic and mechanical stress during tissue dissociation, and the resulting difficulty in obtaining sufficient numbers of viable fibroblasts for downstream experiments. Even small variations in digestion and early culture conditions can affect cell recovery, purity, and expansion, highlighting the need for optimized and reproducible isolation protocols. Here, we compared three enzymatic strategies for establishing primary adult mouse cardiac fibroblast cultures: sequential digestion with collagenase I and trypsin, retrograde coronary perfusion with collagenase II, and continuous digestion with Liberase DH. The three protocols were evaluated by assessing post-isolation cell viability, the composition of recovered non-myocyte populations by flow cytometry, and the ability of isolated cells to establish and expand during the first five days of culture. Retrograde coronary perfusion generated viable fibroblasts but yielded relatively few cells and is most advantageous when fibroblasts are isolated simultaneously with cardiomyocytes from the same heart. Liberase DH digestion recovered a broader spectrum of non-myocyte populations, making it suitable for immediate phenotypic analyses, although it produced less efficient fibroblast expansion in culture. Sequential collagenase I/trypsin digestion showed the best overall performance, with low mortality, negligible CD31⁺ endothelial contamination, and robust expansion into confluent spindle-shaped monolayers within five days. Overall, these findings identify sequential collagenase I/trypsin digestion as an effective and reproducible approach for establishing viable adult mouse cardiac fibroblast cultures for downstream functional and molecular studies. Full article