The incidence of temporomandibular joint (TMJ) degeneration has been steadily increasing, with overloading identified as a major risk factor. This condition often leads to condylar cartilage degeneration, significantly affecting patients' quality of life; however, the molecular mechanisms underlying this process remain poorly understood, and effective treatments are still lacking. We utilized single-nucleus RNA sequencing to analyze the condylar cartilage in an overloading mouse model. This approach enabled the identification of 11 distinct cell types within the condylar chondrocytes. Through the application of pseudotime trajectory Analysis and cellchat analyses, we identified the key gene Acvr1b and its associated signaling pathway, which are crucial for regulating the terminal differentiation of condylar chondrocytes. This study utilized single-nucleus RNA sequencing and in vitro validation to investigate the role of Acvr1b in TMJ cartilage degeneration under overloading stress. Our findings reveal key pathways involved in chondrocyte differentiation, providing a theoretical basis for the development of targeted therapeutic interventions.

• Klíčová slova
• Acvr1b, condylar cartilage, single cell, temporomandibular joint, terminal differentiation,
• Publikační typ
• časopisecké články MeSH

Andrographolide (1) is a labdane-type diterpene lactone and the major bioactive metabolite (2.39%) in the leaves of Andrographis paniculata (Acanthaceae). To further explore its stability, the thermal degradation kinetics of compound 1 at pH 2.0, pH 6.0, and pH 8.0 were modeled at three temperatures within 50-85 °C. The activation energy (Ea), shelf-life (t90%), and rate constant (k) for compound 1 were determined using the Arrhenius equation. Consequently, the results indicated that degradation followed first-order kinetics, and the optimum pH for stability was determined to be between pH 2.0 and 4.0. Major degradation products formed under pH 2.0 and pH 6.0 conditions were isolated and spectroscopically characterized by comparison with known compounds. Under pH 2.0 conditions, two degradation products were identified: isoandrographolide (2) and 8,9-didehydroandrographolide (3). Under pH 6.0 conditions, three degradation products were formed: 15-seco-andrographolide (4), 14-deoxy-15-methoxyandrographolide (5), and 11,14-dehydro-14-deoxyandrographolide (6). Anti-inflammatory and cytotoxicity assessments demonstrated reduced biological effects for the degradation products compared with compound 1. This highlights the importance of controlling pH during formulation to ensure product stability, sustained bioactivity, and patient benefit.

• Klíčová slova
• Andrographolide, Arrhenius equation, Cytotoxicity, Degradation, NO production, Product characterization, pH dependency,
• MeSH
• Andrographis chemie   MeSH
• antiflogistika farmakologie   chemie   MeSH
• diterpeny * chemie   farmakologie   MeSH
• kinetika MeSH
• koncentrace vodíkových iontů MeSH
• lidé MeSH
• listy rostlin chemie   MeSH
• myši MeSH
• stabilita léku MeSH
• teplota MeSH
• voda chemie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• andrographolide MeSH Prohlížeč
• antiflogistika MeSH
• diterpeny * MeSH
• voda MeSH

Brain ischemia is a severe condition caused by reduced cerebral blood flow, leading to the disruption of ion gradients in brain tissue. This imbalance triggers spreading depolarizations, which are waves of neuronal and glial depolarization propagating through the gray matter. Microelectrode arrays (MEAs) are essential for real-time monitoring of these electrophysiological processes both in vivo and in vitro, but their sensitivity and signal quality are critical for accurate detection of extracellular brain activity. In this study, we evaluate the performance of a flexible microelectrode array based on gold-coated zinc oxide nanorods (ZnO NRs), referred to as nano-fMEA, specifically for high-fidelity electrophysiological recording under pathological conditions. Acute mouse brain slices were tested under two ischemic models: oxygen-glucose deprivation (OGD) and hyperkalemia. The nano-fMEA demonstrated significant improvements in event detection rates and in capturing subtle fluctuations in neural signals compared to flat fMEAs. This enhanced performance is primarily attributed to an optimized electrode-tissue interface that reduces impedance and improves charge transfer. These features enabled the nano-fMEA to detect weak or transient electrophysiological events more effectively, making it a valuable platform for investigating neural dynamics during metabolic stress. Overall, the results underscore the promise of ZnO NRs in advancing electrophysiological tools for neuroscience research.

• Klíčová slova
• acute brain slices, cerebral ischemia, micro/nano electrode array, spreading depolarization, zinc oxide nanorods,
• Publikační typ
• časopisecké články MeSH

BACKGROUND: This study investigates the role of the SLC38A8 gene. SLC38A8 facilitates glutamine influx, which converts to glutamate in the visual pathway. Mutations in SLC38A8 are associated with FHONDA syndrome, a subtype of foveal hypoplasia with congenital nystagmus and optic-nerve-decussation defects without pigmentation leading to severe vision loss. METHODS: In vivo and in vitro methods were conducted using retinal cell lines overexpressing SLC38A8, and Slc38a8/Slc38a7 gene-edited mice to evaluate visual function and physiological changes. Statistical analyses included two-way ANOVA, multiple regression, and ANCOVA. RESULTS: In vitro, SLC38A8 overexpression influenced retinal gene expression, light detection, and visual perception, as well as glutamine and glutamate dynamics. In Y79SNAT8-OE cells, glutamate levels were significantly higher under light conditions compared to dark conditions at 12 h (3.4 ± 0.16 nmol/μl vs. 3.9 ± 0.17 nmol/μl, p = 0.0011) and 17 h (3.6 ± 0.22 nmol/μl vs. 4.5 ± 0.24 nmol/μl, p = 0.0001), a pattern not observed in control cells. SLC38A8 expression also increased significantly (RQ = 2.1 ± 0.11, p < 0.05) in Y79 cells under glutamine deprivation. In vivo, Slc38a8-truncated gene mice exhibited altered testicular morphology, with significantly reduced volume (70.9 ± 5.1 mm3 vs. 85.5 ± 6.7 mm3, p = 0.023), and reduced length (4.8 ± 0.2 mm vs. 5.4 ± 0.4 mm, p = 0.0169), alongside degenerative changes in germinal epithelium, and elevated liver enzyme. Despite normal eye morphology, retinal thickness, and visual evoked potentials, electroretinogram and behavioural tests indicated enhanced scotopic responsiveness with significant increases in a-wave (162.98 ± 14.1 μv vs. 133.9 ± 36.9 μv, p = 1.5e-07) and b-wave amplitudes (274.82 ± 25.2 μv vs. 199.9 ± 56.1 μv, p = 3.02e-09). CONCLUSIONS: Our findings underscore SLC38A8 role in retinal function and glutamine-glutamate metabolism, with clinical implications for FHONDA and potential future dietary intervention targeting glutamine or glutamate.

• Klíčová slova
• SLC38A8, FHONDA syndrome, glutamine‐glutamate cycle, phototransduction, retinal function,
• MeSH
• elektroretinografie MeSH
• glutamin metabolismus   MeSH
• kyselina glutamová metabolismus   MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nemoci retiny * genetika   metabolismus   patofyziologie   MeSH
• regulace genové exprese * fyziologie   MeSH
• retina * metabolismus   MeSH
• transportní systém aminokyselin y+ * genetika   MeSH
• zrakové dráhy * metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• glutamin MeSH
• kyselina glutamová MeSH
• transportní systém aminokyselin y+ * MeSH

The Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) infection is not limited to the respiratory tract as receptors, including the angiotensin-converting enzyme 2 (ACE2), are expressed across many tissues. This study employed a new conditional mouse model, Rosa26creERT2/chACE2, which expresses human ACE2 (hACE2) across multiple organs, to investigate the effects of SARS-CoV-2 infection beyond the respiratory system. This strain demonstrated susceptibility to SARS-CoV-2 infection in a dose and sex-dependent manner, showing that infected male mice exhibited more severe disease outcomes, including significant weight loss, pronounced lung pathology and dysfunction, and increased mortality, compared to females. In contrast to intratracheal infection, intranasal virus administration facilitated viral spread to the brain, thereby underscoring the nasal route's role in the pathogenesis of neurological manifestations. Intranasal infection also led to increased innate immune system activation as compared to intratracheal virus administration, even though both routes activated the adaptive immune response. This model provides a valuable tool to study SARS-CoV-2 in individual tissues or use a multisystemic approach, and it also advances possibilities for preclinical evaluation of antiviral therapies and vaccine strategies.

• Klíčová slova
• Conditional mouse model, Infection, SARS-CoV-2, hACE2,
• MeSH
• angiotensin-konvertující enzym 2 * genetika   metabolismus   MeSH
• COVID-19 * patologie   virologie   imunologie   genetika   MeSH
• dýchací soustava virologie   patologie   MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• myši transgenní MeSH
• myši MeSH
• plíce virologie   patologie   MeSH
• přirozená imunita MeSH
• SARS-CoV-2 * patogenita   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ACE2 protein, human MeSH Prohlížeč
• angiotensin-konvertující enzym 2 * MeSH

Histone deacetylases (HDACs) are frequently deregulated in cancer, and several HDAC inhibitors (HDACi) have gained approval for treating peripheral T cell lymphomas. Here, we investigated the effects of pharmacological or genetic HDAC inhibition on NPM::ALK positive anaplastic large cell lymphoma (ALCL) development to assess the potential use of HDACi for the treatment of this disease. Short-term systemic pharmacological inhibition of HDACs using the HDACi Entinostat in a premalignant ALCL mouse model postponed or even abolished lymphoma development, despite high expression of the NPM::ALK fusion oncogene. To further disentangle the effects of systemic HDAC inhibition from thymocyte intrinsic effects, conditional genetic deletions of HDAC1 and HDAC2 enzymes were employed. In sharp contrast, T cell-specific deletion of Hdac1 or Hdac2 in the ALCL mouse model significantly accelerated NPM::ALK-driven lymphomagenesis, with Hdac1 loss having a more pronounced effect. Integration of gene expression and chromatin accessibility data revealed that Hdac1 deletion selectively perturbed cell type-specific transcriptional programs, crucial for T cell differentiation and signaling. Moreover, multiple oncogenic signaling pathways, including PDGFRB signaling, were highly upregulated. Our findings underscore the tumor-suppressive function of HDAC1 and HDAC2 in T cells during ALCL development. Nevertheless, systemic pharmacological inhibition of HDACs could still potentially improve current therapeutic outcomes.

• MeSH
• anaplastická lymfomová kináza * metabolismus   genetika   MeSH
• anaplastický velkobuněčný lymfom * farmakoterapie   patologie   genetika   metabolismus   MeSH
• benzamidy farmakologie   MeSH
• histondeacetylasa 1 * genetika   antagonisté a inhibitory   fyziologie   metabolismus   MeSH
• histondeacetylasa 2 genetika   MeSH
• inhibitory histondeacetylas * farmakologie   terapeutické užití   MeSH
• lidé MeSH
• myši MeSH
• pyridiny farmakologie   MeSH
• tumor supresorové geny * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• Alk protein, mouse MeSH Prohlížeč
• anaplastická lymfomová kináza * MeSH
• benzamidy MeSH
• entinostat MeSH Prohlížeč
• Hdac1 protein, mouse MeSH Prohlížeč
• histondeacetylasa 1 * MeSH
• histondeacetylasa 2 MeSH
• inhibitory histondeacetylas * MeSH
• pyridiny MeSH

Activation of immune response plays an important role in the development of retinal diseases. One of the main populations of immune cells contributing to the retinal homeostasis are microglia, which represent a population of residential macrophages. However, under pathological conditions, microglia become activated and rather support a harmful inflammatory reaction and retinal angiogenesis. Therefore, targeting these cells could provide protection against retinal neuroinflammation and neovascularization. In the recent study, we analyzed effects of silver nanoparticles (AgNPs) on microglia in vitro and in vivo. We showed that the AgNPs interact in vitro with stimulated mouse CD45/CD11b positive cells (microglia/macrophages), decrease their secretion of nitric oxide and vascular endothelial growth factor, and regulate the expression of genes for Iba-1 and interleukin-1β (IL-1β). In our in vivo experimental mouse model, the intravitreal application of a mixture of proinflammatory cytokines tumor necrosis factor-α, IL-1β and interferon-γ induced local inflammation and increased local expression of genes for inducible nitric oxide synthase, IL-α, IL-1β and galectin-3 in the retina. This stimulation of local inflammatory reaction was significantly inhibited by intravitreal administration of AgNPs. The application of AgNPs also decreased the presence of CD11b/Galectin-3 positive cells in neuroinflammatory retina, but did not influence viability of cells and expression of gene for rhodopsin in the retinal tissue. These data indicate that AgNPs regulate reactivity of activated microglia in the diseased retina and thus could provide a beneficial effect for the treatment of several retinal diseases.

• Klíčová slova
• Microglia, Neovascularization, Neuroinflammation, Retinal diseases, Silver nanoparticles,
• MeSH
• kovové nanočástice * aplikace a dávkování   MeSH
• makrofágy účinky léků   imunologie   MeSH
• mikroglie účinky léků   imunologie   metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• retina * účinky léků   imunologie   patologie   MeSH
• stříbro * farmakologie   aplikace a dávkování   terapeutické užití   MeSH
• zánět farmakoterapie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• stříbro * MeSH

BACKGROUND AND AIMS: Hypertrophic cardiomyopathy (HCM) is a genetic cardiac disorder characterized by left ventricular hypertrophy (LVH), diastolic dysfunction, and impaired metabolic efficiency. This study investigates the therapeutic potential of the sodium-glucose cotransporter 2 inhibitor (SGLT2i) empagliflozin (EMPA) in ameliorating these pathological features in a mouse model carrying the myosin R403Q mutation. METHODS: Male mice harbouring the R403Q mutation were treated with EMPA for 16 weeks. Multi-nuclear magnetic resonance spectroscopy (31P, 13C, and 23Na MRS), echocardiography, transcriptomic, proteomic, and phosphoproteomic profiling were utilized to assess metabolic, structural, and functional changes. RESULTS: Empagliflozin facilitated the coupling of glycolysis with glucose oxidation and normalized elevated intracellular sodium levels. Treatment resulted in a significant reduction in LVH and myocardial fibrosis as evidenced by echocardiography and histopathology. These structural improvements correlated with enhancements in mitochondrial adenosine triphosphate (ATP) synthesis, fatty acid oxidation, and branched-chain amino acid catabolism. Furthermore, EMPA improved left ventricular diastolic function and contractile reserve, underscored by improved ATP production and reduced energy cost of contraction. Notably, these benefits were linked to down-regulation of the mammalian target of rapamycin signalling pathway and normalization of myocardial substrate metabolic fluxes. CONCLUSIONS: Empagliflozin significantly mitigates structural and metabolic dysfunctions in a mouse model of HCM, underscoring its potential as a therapeutic agent for managing this condition. These findings suggest broader applicability of SGLT2i in cardiovascular diseases, including those due to myocardial-specific mutations, warranting further clinical investigation.

• Klíčová slova
• Branched-chain amino acids, Cardiac energetics, Cardiac metabolism, Hypertrophic cardiomyopathy, Metabolic reprogramming, SGLT2 inhibition, Uncoupled glycolysis, mTOR,
• Publikační typ
• časopisecké články MeSH

Sex-specific penetrance in autosomal dominant Mendelian conditions is largely understudied. The neurodevelopmental disorder Pilarowski-Bjornsson syndrome (PILBOS) was initially described in females. Here, we describe the clinical and genetic characteristics of the largest PILBOS cohort to date, showing that both sexes can exhibit PILBOS features, although males are overrepresented. A mouse model carrying a human-derived Chd1 missense variant (Chd1 R616Q/+) displays female-restricted phenotypes, including growth deficiency, anxiety and hypotonia. Orchiectomy unmasks a growth deficiency phenotype in male Chd1 R616Q/+ mice, while testosterone rescues the phenotype in females, implicating androgens in phenotype modulation. In the gnomAD and UK Biobank databases, rare missense variants in CHD1 are overrepresented in males, supporting a male protective effect. We identify 33 additional highly constrained autosomal genes with missense variant overrepresentation in males. Our results support androgen-regulated sexual dimorphism in PILBOS and open novel avenues to understand the mechanistic basis of sexual dimorphism in other autosomal Mendelian disorders.

• Klíčová slova
• CHD1, Mendelian disease, Neurodevelopmental disorder, Sex differences,
• Publikační typ
• časopisecké články MeSH
• preprinty MeSH

To address the challenge of drug accumulation and penetration at the tumor site(s), herein we describe a first-in-class nanocarrier containing 24 copies each of two bioactive peptides (BAPs) genetically fused in frame to the 24 N-termini of a human ferritin H-type construct, named THE-10. The two BAPs are specific for PD-L1 and integrin αVβ3/αVβ5 plus Neuropilin (iRGD) respectively, conferring immune checkpoint blockade and drug-internalization properties. In turn, the THE-10 backbone brings 48 BAPs contiguous for synergism, prolonged blood half-life, and release into the tumor microenvironment upon conditional cleavage of a metalloprotease-sensitive site. Predicted THE-10 multitasking activity was experimentally supported as follows. Size-exclusion chromatography and surface plasmon resonance demonstrated BAP cleavage/release and receptor binding (nanomolar KD). Live-cell/time-lapse imaging demonstrated 4-fold-increased internalization of naked therapeutic antibodies, mirrored by enhanced cytotoxicity of the corresponding Antibody-Drug Conjugate. Slight antitumor effects were observed in vivo by treating immune checkpoint-sensitive syngeneic mouse colorectal model with THE-10 alone. Drug boosting was instead considerable on colorectal and pancreatic tumor allografts when THE-10 was co-administered with both small and large chemotherapeutic agents, outperforming the original iRGD cyclic peptide. Thus, THE-10 may enhance target therapy, chemotherapy and immunotherapy altogether, e.g. it candidates as a multitasking, all-round, antineoplastic therapy booster.

• Klíčová slova
• Human ferritin nanocarrier, Peptide grafting, cancer therapy booster,
• MeSH
• ferritiny * chemie   genetika   farmakologie   MeSH
• imunoterapie * MeSH
• lidé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nanočástice * chemie   MeSH
• nosiče léků * chemie   MeSH
• protinádorové látky farmakologie   chemie   MeSH
• rekombinantní proteiny chemie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ferritiny * MeSH
• nosiče léků * MeSH
• protinádorové látky MeSH
• rekombinantní proteiny MeSH