Search Results (26,143)

Both RIT1 and RIT2 are members of the RAS superfamily of small GTPases, which regulate various cellular processes. RIT1 is widely expressed, whereas RIT2 is primarily found in neuronal tissues. Dysregulation of these proteins has been associated with several human diseases, including Noonan syndrome, cancer, Parkinson’s disease, autism, and schizophrenia. Although RIT1 and RIT2 are often compared to classical RAS proteins, they exhibit distinct regulatory and biochemical properties. Here, we demonstrate that RIT1 differs from classical RAS in GTPase cycling. Unlike classical RAS proteins, RIT1 did not respond to SOS1-mediated nucleotide exchange or p120GAP-stimulated GTP hydrolysis under cell-free conditions. These results imply that RIT1 may depend on regulatory mechanisms that differ from those of classical RAS proteins. However, the relevant physiological regulators remain unknown. Disease-associated RIT1 mutations cluster around the P-loop and Switch II regions. In this transient overexpression screening system, however, these mutations had only a modest effect on the canonical MAPK, PI3K/AKT, and JNK signaling pathways in HEK293T overexpression experiments. This suggests the existence of additional context-specific effectors and regulatory factors. We demonstrate that RIT1 and RIT2 interact with membrane lipids via a basic C-terminal extension. The KRLK-containing region contributes to the binding of phosphatidylserine and phosphoinositides. Charge-reversal mutations disrupt lipid interactions and liposome binding, supporting the functional importance of this region. In a reconstituted liposome system, galectin-3 and LZTR1, but not galectin-1, reduced the interaction of GDP-loaded RIT1 and RIT2 with liposomes. These results suggest that accessory proteins may influence RIT membrane interactions. However, their cellular relevance requires further validation. Together, our findings provide biochemical insights into RIT GTPase regulation and its interactions with membrane lipids under cell-free conditions. Full article

Fusarium mycotoxins are major contaminants of human and animal feed and pose significant risks to health. A recent recommendation from the European Commission drastically decreased the maximum tolerated concentrations of fumonisins (FBs) and deoxynivalenol (DON) in poultry feed. The objectives of this study were to characterize the effects of a 14-day exposure to a diet containing 7.5 mg FB1 + FB2/kg, 2.5 mg DON/kg and 0.6 mg zearalenone (FDZ diet) in turkeys and chickens, and to assess the persistence of these effects following withdrawal of the contaminated diet. No clinical signs of toxicity were observed in either species, and markers of hepatic oxidative damage remained unchanged. In contrast, sphingolipidome alterations were detected, being more pronounced in turkeys than chickens and more persistent in the liver than in plasma. The C22–C24:C16 ratios measured across several sphingolipid (SL) classes emerged as sensitive biomarkers of FB exposure, whereas the sphinganine:sphingosine ratio (Sa:So) remained unchanged. Determination of the 90th percentile (P90) thresholds for SL ratios in unexposed animals provided good-to-excellent discrimination between exposed and unexposed animals. Notably, this conservative percentile-based approach detected FB exposure using the Sa:So ratio despite the absence of significant differences in group means. Multivariate analysis of the complete sphingolipidome provided the highest discriminatory power, distinguishing exposed from unexposed turkeys and chickens for up to four days after withdrawal of the FDZ diet. Finally, the biological significance of alterations in d18:1P, 18:0/2:0, 18:1/16:0, and dihydrosphingolipids induced by the FDZ diet is discussed. Full article

The posterior communicating artery (PComA) aneurysm is a challenging microsurgical problem due to multiple factors. These include the technical complexity of the surgery itself, potential injury to blood vessels during surgery, recovery of cranial nerve function, and overall neurological outcome after the operation. These are all related to the area where the PComA aneurysm is located, but they have fundamentally different biological determinants. Prior methods of describing aneurysms do not adequately describe how the relationships of the internal carotid artery (ICA) and PComA/P1 configuration influence the proximity of the aneurysm to other important structures such as the perforating arteries, the anterior choroidal artery (AChA), cranial nerve III (CN III), and the surgical corridor. We created PComA-CORE, an artificial intelligence-based framework designed to evaluate whether the elements of experienced microsurgeons’ thought processes can be measured individually while still maintaining temporally valid predictions, human interpretability, and explicit estimates of predictive uncertainty. Methods: Using a highly detailed database of clinical, radiographic, anatomical, intraoperative, and longitudinal data from 687 adult patients who underwent microsurgical clipping of PComA aneurysms over the period 1997–2026, we applied PComA-CORE to predict separately: Microsurgical Complexity (C); Oculomotor Recovery (O); Neurovascular Preservation Risk (R); and Expected 90-Day Functional Outcome (E). The models used cases from 1997–2020 (n = 564) for development and cases from 2021–2026 (n = 123) for temporal evaluation. Several architectures, including penalized regression, machine-learning techniques, interpretable machine learning, and ensembles, were compared using nested cross-validation, discrimination metrics, calibration metrics, decision-curve analysis, explainability measures, uncertainty-aware prediction, inter-observer reproducibility, and model-to-score distillation. Results: Four discrete predictive architectures were identified by PComA-CORE. PComA-C was found to be highly dependent upon anatomy because the specific geometric characteristics of individual vascular segments and the presence or incorporation of branches around the aneurysm strongly influenced temporal predictions. PComA-R was found to behave as a distributed susceptibility phenotype based on neurovascular attributes rather than a deterministic injury model and achieved a temporal AUC of 0.703. Among patients with preoperative CN III palsy, PComA-O identified that recovery primarily depended upon the time course of neurological dysfunction and structural deformation of the affected nerve. Temporal validation was not feasible given the small number of recent non-recovery events. Conversely, PComA-E showed that global functional outcome continued to depend predominantly upon clinical neurological severity, with a temporally evaluated penalized model achieving an AUC of 0.878. Uncertainty-aware predictions indicated that some cases would benefit from greater caution in interpretation. High-resolution anatomical phenotypes demonstrated good inter-observer reproducibility. Score distillation demonstrated that simplification preserved predictive information, but did so differently depending on the endpoint. Conclusions: The problem of predicting the consequences of clipping a PComA aneurysm is multidimensional and does not exist as a single “risk” prediction problem. Technical complexity, neurovascular vulnerability, neural recovery, and global disability each exist within distinct predictive spaces and require different levels of anatomical detail and/or computational complexity. PComA-CORE establishes a human-supervised framework to transform expert microsurgical thought processes into explicit, reproducible, uncertainty-aware, and clinically interpretable representations. While prospective multicenter validation will be needed prior to clinical use, it has the potential to establish a basis for explainable AI, precision cerebrovascular neurosurgery, anatomy-informed risk stratification, and clinically interpretable decision-support systems in complex aneurysm surgery. Full article

Background: Building on our recent characterization of a layer-specific neurovascular microenvironment in the normal human vocal fold, we examined how that unit behaves in laryngo-hypopharyngeal carcinoma, where lymphatic and venous involvement are conventionally reported together. Methods: We retrospectively analyzed 48 laryngo-hypopharyngeal squamous cell carcinomas (2017–2025) with complete TNM vascular reporting, confirmed by CD31 and neuron-specific enolase (NSE) immunohistochemistry. Lymphatic (L) versus venous (V) assignment rested on morphological criteria, since CD31 cannot discriminate lymphatic from blood vessels and D2-40 was unavailable. L, V and perineural (Pn) status were related to pT stage and topography and combined into a composite score (NV = L + V + Pn), reassessed after adjustment for pT. Results: Concordant patterns predominated (L0V0 60.4%, L1V1 27.1%); 12.5% were dissociated. Vascular invasion correlated with pT (ρ = 0.393, p = 0.006) and, unadjusted, with Pn, but not after adjustment for pT. NV tracked pT more strongly than any single axis (KR-20 = 0.725; ρ = 0.534, p < 0.001). Glottic-only carcinoma showed the lowest involvement on all axes. Conclusions: Reporting L, V and Pn separately—parameters already in routine TNM notation—may capture tumor aggressiveness more completely than LVI as a single entity. This series cannot establish stage-independent co-variation in these axes, validate the L/V distinction molecularly, or, lacking follow-up, demonstrate prognostic value. Full article

Accurate river water quality assessment requires precise knowledge of human activities in the catchment. The water quality of the Katse Reservoir inflow rivers (KRIRs) is affected by mining effluents and livestock waste to varying degrees. This study compared the cumulative impact of human activities on the KRIRs through a correlation analysis and agglomerative hierarchical clustering of the water quality index (WQI) and the eutrophication index (EI) during the first decade (FD) (2003 to 2013) and the second decade (SD) (2014 to 2024). The study found that the water quality parameters selected to compute the WQI (NO₃⁻, PO₄³⁻, NH₄⁺, NO₂⁻, Fe²⁺, Cd²⁺, Pb²⁺, EC, and pH) fluctuated within the water quality standards except for chemical oxygen demand (COD) and Escherichia coli (E. coli). Dissolved inorganic nitrogen, dissolved inorganic phosphorus, dissolved oxygen, and COD were used to compute the EI. The results from all water quality sites (WQS) showed that water was unsuitable for domestic use (WQI < 50). WQS-E and WQS-D were suitable for industrial use (51 < WQI < 75), whereas WQS-A, WQS-B and WQS-C were unsuitable for any use before treatment (100 < WQI). E. coli contributed significantly to the WQI, with strong correlations in the FD (r = 0.98) and in the SD (r = 0.94) (p < 0.023). Nitrates contributed significantly to the EI in the SD (r = 0.67). The correlation between WQI and EI increased from r = 0.45 in the FD to r = 0.81 in the SD at a significance level of α = 0.05. The study effectively highlighted the evolution of pollution from diffuse to point sources in the SD at WQS-A. These findings can be used to forecast water quality responses to future land use activities and guide management strategies to curb pollution at source. Full article

PIK3CA, which encodes the p110α catalytic subunit of phosphoinositide 3-kinase (PI3K), is one of the most frequently altered oncogenes in human cancer and a major driver of tumor initiation, progression, metastasis, and therapeutic resistance. Over the past two decades, advances in structural biology, cancer genomics, and translational research have substantially expanded our understanding of PIK3CA function and established the PI3K pathway as a clinically actionable therapeutic target. This review provides an overview of the structural organization and physiological functions of the PI3Kα complex, the molecular mechanisms underlying oncogenic activation, and the diverse spectrum of PIK3CA alterations across human malignancies. We also summarize the current landscape of PI3K-targeted therapies, highlighting both approved agents and emerging therapeutic strategies. Clinical evidence supports the rational integration of PI3K inhibitors with endocrine therapy, CDK4/6 inhibitors, MAPK pathway inhibitors, dual PI3K/mTOR inhibition, and immune checkpoint blockade. In addition, accumulating evidence indicates that PIK3CA plays a pivotal role in shaping the tumor immune microenvironment, providing a biological rationale for combining PI3K inhibition with immunotherapy. Finally, we discuss future directions in precision oncology, emphasizing integrated molecular profiling, liquid biopsy, single-cell and spatial technologies, functional genomics, and evolutionary approaches as complementary strategies to refine patient selection, overcome therapeutic resistance, and optimize clinical outcomes. Full article

Topical non-steroidal anti-inflammatory drugs (NSAIDs) such as flurbiprofen and benzydamine may be used for the symptomatic relief of pharyngitis, triggered by infections. The onset and spectrum of anti-inflammatory activity of both drugs were compared in human tonsil (HTEpiC) and bronchial epithelial (BEAS-2B) cells stimulated with poly I:C against key inflammatory mediators. Cells were stimulated with poly I:C for 24 h and treated with flurbiprofen or benzydamine for 20 s, 2 min and 5 min. Production of prostaglandin E2 (PGE₂), tumour necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-1 beta (IL-1β), interleukin-8 (IL-8), interleukin-18 (IL-18), monocyte chemotactic protein-3 (MCP-3) and C-X-C motif chemokine ligand 10 (CXCL10), as well as cyclooxygenase-2 (COX-2) protein expression, was evaluated. Flurbiprofen significantly (p < 0.05) reduced PGE₂ production and COX-2 expression in both cell types within 20 s of treatment and maintained these effects at 2 and 5 min. However, benzydamine produced delayed, inconsistent inhibition. Although both drugs reduced poly I:C induced IL-6, IL-1β, IL-8 and IL-18 production, the magnitude and onset were greater with flurbiprofen. Flurbiprofen also reduced MCP-3 and CXCL10 in both cell types, alongside significant inhibition of caspase-1 activity. Benzydamine showed limited or no effect and less inhibition of caspase-1. With rapid suppression of the COX-2/PGE₂ pathway, rapid onset and a broad inhibition spectrum, flurbiprofen more comprehensively modulates inflammatory processes in epithelial respiratory models. Full article

Ticks are hematophagous arthropods that parasitize livestock, humans, and wild animals, posing serious threats to public health worldwide. Diverse pathogens have been discovered in ticks, including viruses, bacteria, protozoa, and filarial nematodes; however, ticks also harbor a variety of fungal microbes, which remain largely unexplored. In this study, we isolated and identified culturable fungi associated with ticks collected on cattle and goats in Yunnan Province, China. Of the 1730 ticks obtained, 1690 were identified as Rhipicephalus microplus, and only 40 as Haemaphysalis longicornis via morphology and 16S rRNA gene analysis. A total of 90 fungal strains were isolated from the ticks and identified by morphological examination and ITS sequencing, representing 25 species across 20 genera, with Fusarium being the most prevalent. Among these isolates, Fusarium verticillioides (19 isolates) poses a threat to grains, animals, and humans; Cladosporium cladosporioides (six isolates) and Sarocladium zeae (one isolate) are primarily pathogenic to plants, while Schizophyllum commune (eight isolates), Rhizopus arrhizus (two isolates), Diaporthe phaseolorum (two isolates), and Purpureocillium lilacinum (two isolates) are recognized animal-associated pathogens. The diversity of isolated fungi with documented pathogenicity suggests that ticks may serve as potential vectors for fungal transmission, raising concerns about their role in cross-kingdom disease spread across agriculture and livestock. Furthermore, fungi with entomopathogenic potential, such as P. lilacinum, offer valuable resources for developing biocontrol agents against ticks. Our findings highlight the dual role of ticks as both potential vectors of phytopathogenic fungi and reservoirs of entomopathogenic biocontrol agents. This necessitates a paradigm shift in tick surveillance, integrating mycological monitoring to mitigate agricultural losses and exploring tick-borne fungi for sustainable vector control strategies. Full article

Inula helenium L. (elecampane) is a traditional medicinal plant whose roots contain both hydrophilic polyphenols and lipophilic sesquiterpene lactones. However, the influence of the raw material origin and the extraction solvent on its combined bioactivity profile remains unclear. Moreover, although isolated sesquiterpene lactones from I. helenium have been extensively investigated for their anticancer properties, the bioactivity of whole root extracts against human melanoma cells has remained largely underexplored. This study aimed to systematically compare the phytochemical characteristics and biological activity of I. helenium root extracts obtained from three distinct raw material sources and three solvents of increasing polarity, and to identify the main factors associated with the observed bioactivity through multivariate integration. In this study, I. helenium roots from three distinct sources—commercially available dried roots, wild-collected roots, and cultivated roots collected from the Botanical Garden of Vilnius University—were extracted with three solvents of increasing polarity (n-hexane, ethyl acetate, and 70% ethanol). Each extract was characterized by total phenolic content, four antioxidant assays (DPPH, ABTS, FRAP, CUPRAC), UV–Vis fingerprinting, antimicrobial testing against Staphylococcus aureus, Klebsiella pneumoniae, and Candida albicans, and a CCK-8 assay on SK-MEL-28 human melanoma and HEK293 non-cancerous cells. Multivariate integration (PCA, HCA, Spearman rank correlation) identified solvent polarity as the dominant driver of both the phytochemical and the biological profile (PC1 and PC2 together explained 87.9% of the total variance). Ethanolic extracts combined the highest phenolic content (up to 38.6 mg GAE/g dry extract) and the strongest antioxidant capacity across all four assays, with maximum values of 177.3 (DPPH), 328.9 (ABTS), 174.1 (FRAP), and 541.6 (CUPRAC) µmol TE/g DE (p ≤ 0.004). In the preliminary antimicrobial screening, ethanolic extracts showed near-complete to complete inhibition of S. aureus and C. albicans at 25 mg/mL, with MIC₉₀ values as low as 12.5 mg/mL against S. aureus and 6.25 mg/mL against C. albicans, whereas K. pneumoniae showed only marginal growth inhibition (median values below 20% across the entire tested concentration range). Strikingly, n-hexane and ethyl acetate extracts—despite negligible antioxidant activity—produced concentration-dependent cytotoxic effects. Six of the nine extracts reduced SK-MEL-28 viability while sparing HEK293 cells, with the cultivated Botanical Garden n-hexane extract showing the greatest difference across the tested cell models (SI > 1.50, p = 0.0003). Targeted solvent selection thus enables a single plant material to yield two functionally distinct bioactive fractions: more polar extracts with pronounced antioxidant and antimicrobial activities and less polar extracts with preferential activity against SK-MEL-28 within the cell models tested. Full article

Background: The human amniotic membrane (hAM) is widely used in tissue engineering due to its low immunogenicity and extracellular matrix (ECM) rich in structural proteins; in this context, decellularization aims to remove cellular components while preserving tissue integrity, and this study aimed to compare the effects of two enzymatic methods (trypsin and thermolysin). Methods: Human amniotic membranes obtained from placentas were divided into three groups: non-decellularized control, trypsin-treated, and thermolysin-treated. The membranes were treated with 0.25 trypsin-EDTA for 60 min at 37 °C, followed by incubation with Triton X-100 for 30 min at 37 °C, or with 125 µg/mL thermolysin for 9 min at 37 °C. Histological analysis with hematoxylin–eosin and semi-quantitative scoring assessed epithelial and stromal integrity, immunohistochemistry evaluated laminin α and fibronectin expression with quantitative image analysis, and uniaxial tensile testing measured mechanical properties; no preregistration numbers or animal models were reported. Results: Trypsin achieved more effective epithelial removal, whereas thermolysin caused greater stromal disruption; no statistically significant differences in laminin α or fibronectin expression were found among groups (p > 0.05), indicating preservation of key ECM components; both treatments reduced mechanical performance, with thermolysin decreasing early- and mid-path stiffness and trypsin reducing late-path stress and work density. Conclusion: Both enzymatic protocols preserved ECM composition but induced distinct structural and mechanical alterations, suggesting that method selection should balance efficient cell removal with preservation of structural and functional properties according to the intended application. Full article

This study evaluated and compared four irrigation activation protocols—conventional syringe irrigation (CSI), passive ultrasonic irrigation (PUI), photon-induced photoacoustic streaming (PIPS), and shock wave-enhanced emission photoacoustic streaming (SWEEPS)—with 17% EDTA as a final irrigant, with respect to smear layer (SL) removal and push-out bond strength (PBS) of root canal sealer in the apical third of curved root canals. One hundred and twenty extracted single-rooted human premolars with oval canals and moderate curvature (20–40°, Schneider’s method) were allocated into four groups randomly (n = 30 each). Canals were prepared crown-down under 2.5% NaOCl irrigation, followed by final 17% EDTA irrigation per the assigned protocol: CSI, PUI, PIPS, and SWEEPS. 10 specimens per group underwent SEM evaluation of SL removal using Hülsmann’s criteria; remaining specimens were obturated. In 10 samples, apical sealer adaptation was assessed via SEM. PBS of ten samples were evaluated using a universal testing machine, followed by failure mode assessment. Data were statistically evaluated using two-way mixed ANOVA, followed by Tukey’s post hoc test for PBS and Kruskal–Wallis test with Dunn’s multiple-comparison test with Bonferroni correction for SL scores (p < 0.05). Minimum SL removal was exhibited by the apical third of Group 1 (CSI-EDTA) samples. PIPS-EDTA and SWEEPS-EDTA presented comparable SL elimination outcomes in the cervical and middle thirds (p > 0.05). SL elimination between these two groups was significantly different in the apical third. The cervical section of Group 4 displayed the highest bond integrity. However, the lowest bond strength was exhibited by the apical third of Group 1 (CSI-EDTA). SWEEPS combined with EDTA demonstrated superior smear layer removal throughout curved root canals, including the apical third, and yielded the highest push-out bond strength among all tested irrigation protocols. Full article

Background/Objectives: To compare the diagnostic performance of Claude Opus 4.7 and Gemini Pro 3 for the differential diagnosis of oral autoimmune blistering diseases (AIBDs) and evaluate their diagnostic reasoning, confidence, and calibration. Materials and Methods: This retrospective multicenter paired diagnostic accuracy study included 200 clinicopathologically confirmed AIBD cases (50 each of pemphigus vulgaris, mucous membrane pemphigoid, bullous pemphigoid, and linear IgA bullous dermatosis). Each case was independently assessed by both models using identical standardized clinical information and clinical photographic inputs. The task required forced-choice classification among the four predefined diseases. Histopathological and direct immunofluorescence findings were used exclusively to establish the clinicopathological reference diagnosis and were not provided to the AI models. The reference diagnosis was established by clinicopathological correlation. The primary outcome was diagnostic accuracy. Secondary outcomes included disease-specific diagnostic performance, Cohen’s κ, ROC analysis, calibration, confidence, reasoning quality, management recommendations, and error patterns. Pre-consensus inter-rater reliability of the two human assessors was also evaluated using Cohen’s κ for binary outcomes and weighted Cohen’s κ for the ordinal reasoning-quality score. Results: Claude achieved significantly higher diagnostic accuracy than Gemini (92.0% vs. 86.0%, p = 0.012), stronger agreement with the reference standard (κ = 0.893 vs. 0.813), and superior discrimination (macro-AUC 0.998 vs. 0.965). Claude demonstrated higher key diagnostic-feature identification (92.0% vs. 86.0%; p = 0.012) and higher clinical-reasoning scores (61.0% vs. 40.0% of responses rated good; Wilcoxon p < 0.001; r = 0.47), whereas management recommendations did not differ significantly (100.0% vs. 98.0%; p = 0.125). Calibration results were metric-dependent: Claude had a lower one-vs-rest Brier score (0.0468 vs. 0.0558), whereas Gemini had a lower expected calibration error (0.083 vs. 0.251). For both models, the predominant error was misclassification of linear IgA bullous dermatosis as mucous membrane pemphigoid. Conclusions: Both multimodal LLMs showed high performance in this controlled four-class benchmark, with Claude Opus 4.7 outperforming Gemini Pro 3 in overall accuracy and reasoning quality. However, these findings do not establish autonomous diagnostic capability, clinical effectiveness, or safety. The LABD–MMP misclassification and metric-dependent calibration highlight important limitations. The models should therefore be regarded as investigational adjunctive decision-support tools requiring clinician oversight and diagnostic verification. Prospective external and human-in-the-loop validation is required before clinical implementation. Full article

Chinese hamster ovary (CHO) cells are the principal platform for manufacturing recombinant biopharmaceuticals, but sustained production can exceed the protein-folding capacity of the endoplasmic reticulum (ER). Binding immunoglobulin protein (BiP/GRP78), encoded by Hspa5, is a major ER chaperone that supports folding and modulates unfolded protein response (UPR) signaling, yet increasing BiP abundance has produced beneficial or inhibitory outcomes depending on the recombinant product. Here, we introduced an additional Chinese hamster Hspa5 coding sequence under the constitutive EEF1A1 promoter into a CHO line producing a dulaglutide analogue comprising a modified GLP-1 peptide fused to human IgG4 Fc and compared the resulting stable DUL-BiP population with an empty-vector DUL-Neo control. At constant 37 °C, the two populations showed similar viable cell density and viability trajectories, whereas DUL-BiP cells continued extracellular GLP-1–Fc accumulation for approximately two days after accumulation in DUL-Neo cells had slowed and reached a 40% higher final titer in this experiment. Two independent temperature-shift experiments showed the same direction of the final-titer difference, although the increases were smaller and were accompanied by reduced peak viable cell density. Semi-quantitative immunoblot analysis of samples from the constant-temperature experiment showed a comparatively stable total BiP signal in DUL-BiP cells and a pronounced late increase in DUL-Neo cells, together with temporal changes in cleaved ATF6, eIF2α phosphorylation, and CHOP in both populations. Intracellular GLP-1–Fc did not progressively accumulate. Because the immunoblot time courses were derived from one biological fed-batch experiment, they are interpreted descriptively rather than as independent quantitative biological replicates. Stable constitutive expression of host-derived BiP was therefore associated with prolonged late-stage GLP-1–Fc accumulation in the producer population examined. These kinetics are compatible with a model in which the timing of ER chaperone supply influences the maintenance of productive output, while the underlying mechanism and generality to other producer populations or recombinant products remain to be established. Full article

Hepatic stellate cells (HSCs) are central drivers of liver fibrosis, yet their heterogeneity is often overlooked in human liver organoid models. Here, we investigated how HSC-intrinsic variability influences fibrotic phenotypes in multicellular liver organoids using induced pluripotent stem cell-derived HSCs (iHSCs) and primary HSCs (pHSCs). iHSCs derived from two different iPSC lines exhibited distinct differentiation phenotypes towards iHSCs. Consistently, co-culture of the hepatocyte-like cell line HepaRG with these two iHSC populations generated organoids with distinct fibrotic and hepatic features. BJ1-iHSC/HepaRG organoids contained an approximately 2.9-fold higher proportion of CYP3A4-positive cells, whereas Sigma-iHSC/HepaRG organoids showed around a 4-fold higher percentage of Sirius red-positive area. Single-cell RNA analysis further showed that the two iHSC populations differed in cell cycle distribution, HSC-related signatures, extracellular matrix-associated pathways, and matrisome enrichment. Similarly, human pHSCs cultured as organoids with HepaRG showed marked donor-dependent differences at baseline and after single acetaminophen, repeated thioacetamide or transforming growth factor beta treatment. Together, our findings suggest that HSC heterogeneity, whether iPSC-derived or donor-derived, is an important factor in fibrotic output and response variability in human liver organoid models and should be considered in model design, interpretation and comparison between studies. Full article

Pulse-processing plants generate large volumes of screenings, that is, undersized, broken and foreign-matter-contaminated grain that is removed before the material enters the human food chain. Their feeding value for ruminants is still poorly documented. This study characterised the chemical composition, Cornell Net Carbohydrate and Protein System (CNCPS) nitrogen fractions, in vitro ruminal dry matter degradability (IVDMD) and rumen fermentation behaviour of screenings from five pulse species: soybean, dry bean, chickpea, green lentil and red lentil. Six independent batches per species (n = 30) were obtained from processing plants in different regions of Türkiye. Total digestible nutrients (TDNs) and energy values were estimated with NRC (2001) equations, gas production was recorded for 48 h in a semi-automatic modular system and fitted to a logistic model, IVDMD was measured in a Daisy^II incubator and volatile fatty acids (VFAs), pH and NH₃-N were determined after 24 h of incubation. Differences were declared at p < 0.05 throughout. Soybean screenings had the highest crude protein (35.99%), recalculated TDNs (75.26%) and 48 h gas production (65.16 mL per 460 mg of incubated sample), a ranking that largely disappears once gas is expressed per gram of incubated organic matter; chickpea screenings combined the highest starch (50.67%), IVDMD (71.27%) and fractional degradation rate. Dry bean screenings fermented most slowly, with a lag time of 10.46 h. Acetate, propionate and total VFAs did not differ among species. No parent grain was analysed alongside the screenings, so any comparison with clean pulse grain rests on published values for other samples and cultivars. Within that limit, the screenings carry enough protein, calculated energy and rumen-undegradable protein to warrant testing as partial replacements for conventional concentrates in feeding trials. Full article