Explore the vast range of titles available.

Physiologically based kinetic (PBK) modelling offers a mechanistic basis for predicting the pharmaco-/toxicokinetics of compounds and thereby provides critical information for integrating toxicity and exposure data to replace animal testing with in vitro or in silico methods. However, traditional PBK modelling depends on animal and human data, which limits its usefulness for non-animal methods. To address this limitation, high-throughput PBK modelling aims to rely exclusively on in vitro and in silico data for model generation. Here, we evaluate a variety of in silico tools and different strategies to parameterise PBK models with input values from various sources in a high-throughput manner. We gather 2000 + publicly available human in vivo concentration–time profiles of 200 + compounds (IV and oral administration), as well as in silico, in vitro and in vivo determined compound-specific parameters required for the PBK modelling of these compounds. Then, we systematically evaluate all possible PBK model parametrisation strategies in PK-Sim and quantify their prediction accuracy against the collected in vivo concentration–time profiles. Our results show that even simple, generic high-throughput PBK modelling can provide accurate predictions of the pharmacokinetics of most compounds (87% of Cmax and 84% of AUC within tenfold). Nevertheless, we also observe major differences in prediction accuracies between the different parameterisation strategies, as well as between different compounds. Finally, we outline a strategy for high-throughput PBK modelling that relies exclusively on freely available tools. Our findings contribute to a more robust understanding of the reliability of high-throughput PBK modelling, which is essential to establish the confidence necessary for its utilisation in Next-Generation Risk Assessment.

Mesothelioma is one of the most aggressive neoplasms worldwide that has a particularly poor prognosis. We have previously discovered that oxytocin receptors (OXTR) are highly expressed in mesothelioma and that OXTR knockdown significantly decreases the proliferation of mesothelioma cells with high OXTR expression. In this study, we performed quantitative proteomic profiling of two mesothelioma cell lines with high OXTR expression using mass spectrometry to investigate the downstream signals of OXTR in mesothelioma cells. We found that OXTR knockdown significantly downregulated PDZ‐binding kinase (PBK)—a serine/threonine protein kinase belonging to the bispecific MAPKK family. PBK knockdown significantly suppressed proliferation, migration, and colony formation in mesothelioma cells with high PBK expression and decreased Akt and MAPK phosphorylation levels. Furthermore, immunohistochemical analysis of PBK in surgical specimens obtained from patients with mesothelioma showed that high PBK expression was strongly associated with poor overall survival (log‐rank test p = 0.0031; hazard ratio 3.339 and 95% confidence interval 1.12–10.00) and recurrence‐free survival (log‐rank test p = 0.0024; hazard ratio 3.355 and 95% confidence interval 1.25–9.04). In addition, the clinical significance of PBK expression was validated in mesothelioma using datasets from TCGA. Multivariable Cox regression analysis, incorporating stage and OXTR mRNA expression, demonstrated that PBK mRNA expression was the strongest independent predictor of OS. Our findings indicate that PBK plays a crucial role in the aggressiveness of mesothelioma, making it a promising therapeutic target and potential prognostic biomarker for mesothelioma. PBK plays a crucial role in cell survival in mesothelioma and is associated with worse outcome. PBK is a promising therapeutic target that may serve as a prognostic biomarker, enabling personalized treatment of mesothelioma.

With an increasing need to incorporate new approach methodologies (NAMs) in chemical risk assessment and the concomitant need to phase out animal testing, the interpretation of in vitro assay readouts for quantitative hazard characterisation becomes more important. Physiologically based kinetic (PBK) models, which simulate the fate of chemicals in tissues of the body, play an essential role in extrapolating in vitro effect concentrations to in vivo bioequivalent exposures. As PBK-based testing approaches evolve, it will become essential to standardise PBK modelling approaches towards a consensus approach that can be used in quantitative in vitro-to-in vivo extrapolation (QIVIVE) studies for regulatory chemical risk assessment based on in vitro assays. Based on results of an ECETOC expert workshop, steps are recommended that can improve regulatory adoption: (1) define context and implementation, taking into consideration model complexity for building fit-for-purpose PBK models, (2) harmonise physiological input parameters and their distribution and define criteria for quality chemical-specific parameters, especially in the absence of in vivo data, (3) apply Good Modelling Practices (GMP) to achieve transparency and design a stepwise approach for PBK model development for risk assessors, (4) evaluate model predictions using alternatives to in vivo PK data including read-across approaches, (5) use case studies to facilitate discussions between modellers and regulators of chemical risk assessment. Proof-of-concepts of generic PBK modelling approaches are published in the scientific literature at an increasing rate. Working on the previously proposed steps is, therefore, needed to gain confidence in PBK modelling approaches for regulatory use.

Background: PDZ-binding kinase/T-LAK cell-originated protein kinase (PBK/TOPK) is a serine–threonine kinase and overexpressed in various types of cancer by inhibiting the transactivation activities of p53 and PTEN. We tested whether PBK/TOPK acts as a cancer-promoting gene through its activation/overexpression in gastric cancer (GC). Methods: We analysed five GC cell lines and 144 primary tumours, which were curatively resected in our hospital between 2001 and 2003. Results: Overexpression of the PBK/TOPK protein was frequently detected in GC cell lines (4 out of 5 lines, 80.0%) was detected in primary tumour samples of GC (24 out of 144 cases, 16.6%) and was significantly correlated with venous invasion, tumour depth and recurrence rate. PDZ-binding kinase/T-LAK cell-originated protein kinase-overexpressing tumours had a worse survival rate than those with non-expressing tumours ( P =0.0009, log-rank test). PDZ-binding kinase/T-LAK cell-originated protein kinase positivity was independently associated with a worse outcome in multivariate analysis ( P <0.0001, hazard ratio 6.40 (2.71–14.49)). In PBK/TOPK-overexpressing GC cells, knockdown of PBK/TOPK inhibited the cell proliferation through the p53 activation in a TP53 mutation-dependent manner and inhibited the migration/invasion through the PTEN upregulation in a TP53 mutation-independent manner. Conclusions: These findings suggest PBK/TOPK plays a crucial role in tumour malignant potential through its overexpression and highlight its usefulness as a prognostic factor and potential therapeutic target in GC.

The human gut microbiota influences host physiology by metabolizing xenobiotics, such as drugs, dietary additives, and environmental contaminants. To enable standardized assessment of microbial xenobiotic metabolism, we established a defined bacterial colon model from pooled human fecal samples, by preparing clean bacterial cell suspensions using density gradient centrifugation. The bacterial cell fraction remained structurally intact, allowing long-term preservation at −70 °C with glycerol as a cryoprotectant. The bacteria were fully reactivated without compromising cellular integrity. Glycerol as a disturbing substrate was removed by washing steps. The system preserved microbial diversity, enzymatic activity, and metabolic functionality over 24 h under anaerobic conditions. Unlike fecal-based models, the model was free from non-cellular contaminants, which could cause unpredictable interactions between test compounds and reactive components within the fecal matrix. Enzymatic assays demonstrated hydrolytic, reductive, and proteolytic activities comparable to native feces. 16S rRNA gene sequencing confirmed taxonomic stability and compositional shifts in response to different substrates. A fiber-rich substrate proved to be optimal for maintaining the bacterial composition over 24 h. We further applied the model to investigate the microbial biotransformation of selected model xenobiotics using high-performance liquid chromatography and mass spectrometry, capturing substrate-specific conversion patterns and formation of expected primary products. A versatile addition of food ingredients, dietary supplements, pharmaceuticals, and environmental chemicals is possible to analyze their effects on the composition of the microbiota, its enzymatic activity, and the excretion of metabolites and end products. Together, this bacterial colon model provides a reproducible, high-throughput capable and ethically viable platform for pharmaco microbiomic studies and next-generation risk assessment. Key points A novel bacterial colon model preserves human gut microbial diversity and activity. Fiber-rich substrates restore enzymatic function and SCFA production. The model enables standardized screening of microbial xenobiotic metabolism.

PDZ-binding kinase/T-lymphokine-activated killer-cell-originated protein kinase (PBK/TOPK) is a serine-threonine protein kinase. This study aimed to investigate the expression pattern, prognostic significance, and relationship with tumor immune infiltration of PBK/TOPK in breast cancer through bioinformatics analyses and immunohistochemistry (IHC), providing insights for individualized treatment and immunotherapy strategies. PBK/TOPK expression and its correlation with clinicopathological features were analyzed using TIMER, UALCAN, and TCGA databases. Prognostic value was assessed via Kaplan–Meier plotter and bc-GenExMiner. Variants of the PBK/TOPK gene and their prognostic implications were explored using cBioPortal. Enrichment analysis in R identified signaling pathways linked to PBK/TOPK. The relationships between PBK/TOPK, tumor-infiltrating lymphocytes (TILs), and immune cell markers were evaluated using TIMER and TISIDB. IHC experiments validated PBK/TOPK expression in tumor samples and its association with prognosis. Meanwhile, IHC also further confirmed the correlation between PBK/TOPK and CD4 + and CD8 + T cells infiltration, as well as the relationship between T cells and breast cancer prognosis. PBK/TOPK was significantly overexpressed in breast cancer tissues compared to normal tissues and correlated with clinicopathological features, including tumor size and lymph node stage. High expression of PBK/TOPK was associated with poor prognosis. While breast cancer subtypes exhibited different PBK/TOPK gene variants, these variants did not influence prognosis. PBK/TOPK is involved in cell meiosis, cytoskeletal motility, and pathways such as FoxO and p53. It is associated with immune infiltration and is differentially expressed in breast cancer subtypes and correlates with TILs and immune cell markers. IHC confirms that PBK/TOPK expression positively correlates with CD4 +  T and CD8 + T cells infiltration. CD4 + T cells were positively associated with the prognosis of triple-negative breast cancer. PBK/TOPK is overexpressed in breast cancer tissues, and high expression levels are associated with poor patient prognosis. Additionally, PBK/TOPK is linked to immune infiltration in breast cancer, positively correlating with CD4 + and CD8 + T cells infiltration, suggesting a new avenue for immunotherapy research in breast cancer.

Children with Group 3 medulloblastoma (G3 MB) have a very poor prognosis, and many do not survive beyond 5 years after diagnosis. A factor that may contribute to this is the lack of available targeted therapy. Expression of protein lin‐28 homolog B ( LIN28B ), a regulator of developmental timing, is upregulated in several cancers, including G3 MB, and is associated with worse survival in this disease. Here, we investigate the role of the LIN28B pathway in G3 MB and demonstrate that the LIN28B–lethal‐7 (let‐7; a microRNA that is a tumor suppressor)–lymphokine‐activated killer T‐cell‐originated protein kinase (PBK; also known as PDZ‐binding kinase) axis promotes G3 MB proliferation. LIN28B knockdown in G3‐MB‐patient‐derived cell lines leads to a significant reduction in cell viability and proliferation in vitro and in prolonged survival of mice with orthotopic tumors. The LIN28 inhibitor N ‐methyl‐ N ‐[3‐(3‐methyl‐1,2,4‐triazolo[4,3‐b]pyridazin‐6‐yl)phenyl]acetamide (1632) significantly reduces G3 MB cell growth and demonstrates efficacy in reducing tumor growth in mouse xenograft models. Inhibiting PBK using HI‐TOPK‐032 also results in a significant reduction in G3 MB cell viability and proliferation. Together, these results highlight a critical role for the LIN28B–let‐7–PBK pathway in G3 MB and provide preliminary preclinical results for drugs targeting this pathway.

The aim of the experiment was to evaluate the process of neutrophil extracellular traps (NETs) formation in patients with oral squamous cell carcinoma (OSCC) in response to direct or indirect contact with SCC cells in comparison to results obtained in the cells of healthy subjects. To fulfill study objectives CAL 27 cell line and blood were obtained from cancer patients and control subjects. Parameters related to NETs formation were analyzed utilizing flow cytometry, fluorescence microscopy, and ELISA-type tests. The expression of selected phosphorylated proteins of the PI3K/Akt/PBK pathway in neutrophils was evaluated using the Western blot method. An increase in NETs formation was observed in a coculture of neutrophils with SCC cells, with the largest amount of NETs formed after stimulation with a supernatant obtained from the SCC culture. The enhanced process of NETs formation was accompanied by changes in the expression of proteins from the PI3K/Akt/PBK pathway. The obtained results prove the existence of interactions between neutrophils and cancer cells resulting in NETosis with the participation of the PI3K/Akt/PBK pathway in patients with OSCC.

Accumulating evidence supports the role of PDZ-binding kinase (PBK)/T-lymphokine-activated killer-cell-originated protein kinase (TOPK) in mitosis and cell-cycle progression of mitotically active cells, especially proliferative malignant cells. PBK/TOPK was confirmed to be associated with the development, progression, and metastasis of malignancies. Therefore, it is a potential therapeutic target in cancer therapy. Many studies have been conducted to explore the clinical applicability of potent PBK/TOPK inhibitors. However, PBK/TOPK has also been shown to be overexpressed in normal proliferative cells, including sperm and neural precursor cells in the subventricular zone of the adult brain, as well as under pathological conditions, such as ischemic tissues, including the heart, brain, and kidney, and plays important roles in their physiological functions, including proliferation and self-renewal. Thus, more research is warranted to further our understanding of PBK/TOPK inhibitors before we can consider their applicability in clinical practice. In this study, we first review the findings, general features, and signaling mechanisms involved in the regulation of mitosis and cell cycle. We then review the functions of PBK/TOPK in pathological conditions, including tumors and ischemic conditions in the heart, brain, and kidney. Finally, we summarize the advances in potent and selective inhibitors and describe the potential use of PBK/TOPK inhibitors in clinical settings.