The health benefits of soy and soy derived foods are often attributed to their isoflavone (ISFs) content, known to include phytoestrogens. However, EFSA has concluded that there is insufficient scientific evidence to establish a causal relationship between ISFs consumption and the proposed health claims. Although phase-II metabolites of ISFs are often considered as detoxified products with reduced estrogenic effects, recent studies indicate that ISFs metabolites may contribute and even enhance the biological activity of ISFs. However, current knowledge on phase-II metabolites of ISFs is limited, even though representing the major ISFs metabolites circulating in the bloodstreams in humans. This study investigates the role of phase-II metabolism in the biological activity of ISFs, focusing on estrogenicity and cytotoxicity in estrogen-sensitive Ishikawa cells (expressing estrogen receptors (ER) α and β). ISFs concentrations from 0.001 to 10 µM, with 1:10 dilution steps, were tested. Cleavage to parent compounds during incubation was also evaluated using high-performance liquid chromatography tandem mass spectrometry. The results showed that phase-II metabolites, particularly sulfates, can still exhibit estrogenic activity, challenging the general assumption that phase-II metabolism always leads to the inactivation of ISFs. In particular, at 10 µM daidzein-4'-sulfate exhibited a comparable estrogenic activity to daidzein at the same concentration, and genistein-7-sulfate exhibited estrogenic potential at higher concentrations. Molecular dynamics simulations revealed that genistein and its conjugates can stably interact with ERβ. This study highlights the importance of phase-II metabolism for ISFs activity and suggests that the sulfates formed should be examined more closely in further investigations.
Since its discovery, the microbiota has been increasingly recognised for its role in maintaining health and contributing to various disease conditions. In the reproductive tract, microbial populations can significantly influence endometrial health, internal homeostasis, and fertility. The preservation or restoration of a balanced microbiota through appropriate probiotic products may support reproductive health; in addition, candidate probiotics must be demonstrated to be safe for use. The aim of this study was to assess the local tolerance and systemic safety of a novel intravaginal probiotic product in cows. Twenty-four animals were enrolled and assigned to four groups: single-dose, three-, and five-fold dose of the test product, and a placebo group receiving excipients only. Physical examination and evaluation of the vaginal mucosa were conducted prior to each treatment, one day after and one and two weeks after the final probiotic administration. Blood and urine samples were collected before treatment and following treatment: one day after the last administration of intravaginal probiotic and again two weeks post-treatment. Blood haptoglobin, serum amyloid A, glucose, non-esterified fatty acids, and beta-hydroxybutyrate were measured to assess inflammatory and metabolic responses, and differences over time and between groups were statistically analysed. No dose-dependent systemic changes were observed; however, transient, time-related alterations were noted across all groups, including controls. The investigational probiotic product was well tolerated both locally and systemically, with tolerability comparable across all groups, including the placebo. These data establish a safety profile in healthy cows and support further investigation of this product in studies focusing on efficacy and microbiome modulation.
High fructose intake has been linked to metabolic and cognitive disturbances, yet its effects on hippocampal synaptic architecture remain unclear. We examined whether four weeks of fructose feeding alter metabolic parameters or CA1 synaptic ultrastructure in adult rats maintained on isocaloric AIN-93G diets containing fructose, glucose, or starch as the primary carbohydrate source. Serum biochemical and hormonal profiles showed only modest, diet-specific differences without major metabolic disruption. Quantitative electron microscopy revealed similar dendritic spine density, postsynaptic density length, perforated synapse frequency, and multisynaptic bouton density across groups, whereas fructose-fed rats displayed a small but significant reduction in spine area and an alteration in circularity. These localized geometric changes occurred without broader synaptic remodeling. Overall, our findings indicate that short-term fructose exposure under metabolically controlled, solid-diet conditions produces minimal metabolic and ultrastructural effects, in contrast to the pronounced disturbances reported in metabolically stressful paradigms, suggesting that structural consequences of fructose depend strongly on dietary context and metabolic load.
Atypical mitosis marks a deviation in the cell division process that has been shown be an independent prognostic marker for tumor malignancy. However, atypical mitosis classification remains challenging due to low prevalence, at times subtle morphological differences from normal mitotic figures, low inter-rater agreement among pathologists, and class imbalance in datasets. Building on the Atypical Mitosis dataset for Breast Cancer (AMi-Br), this study presents a comprehensive benchmark comparing deep learning approaches for automated atypical mitotic figure (AMF) classification, including end-to-end fine-tuned deep learning models, foundation models with linear probing, and foundation models fine-tuned with low-rank adaptation (LoRA). For rigorous evaluation, we further introduce two new held-out AMF datasets - AtNorM-Br, a dataset of mitotic figures from the TCGA breast cancer cohort, and AtNorM-MD, a multi-domain dataset of mitotic figures from a subset of the MIDOG++ training set. We found average balanced accuracy values of up to 0.8135, 0.7788, and 0.7723 on the in-domain AMi-Br and the out-of-domain AtNorm-Br and AtNorM-MD datasets, respectively. Our work shows that atypical mitotic figure classification, while being a challenging problem, can be effectively addressed through the use of recent advances in transfer learning and model fine-tuning techniques. We make all code and data used in this paper available in this github repository: https://github.com/DeepMicroscopy/AMi-Br_Benchmark
The historic Rockborn strain of the canine distemper virus was widely used as a vaccine, but its use was discontinued due to safety concerns. Yet, Rockborn-like canine distemper virus strains are still used in some vaccine formulations. Genetic analysis of this strain was previously limited to the H gene, leaving its full evolutionary and pathogenic potential unclear. This study aimed to determine the complete genome sequence of the Rockborn strain to reconstruct its origin, understand its evolution, and provide a reference for improving diagnostics and future research on virulence markers. An amplicon-based sequencing protocol using MinION nanopore technology was employed to determine the complete genome of the Rockborn-46th laboratory strain. The genome was assembled, annotated, and analyzed in comparison with 223 genomes. The complete genome of the Rockborn strain was 15,690 nucleotides in length. Phylogenetic analysis revealed that Rockborn forms a unique lineage with field isolates from a masked civet in China and a dog in the United States. Crucially, a significant recombination event was identified, showing that the Rockborn strain acted as a parental strain, contributing its F and H genes to create mosaic viruses. The full-genome characterization of the Rockborn strain confirms that Rockborn-like viruses persist and actively contribute to the evolution of canine distemper virus through recombination. This finding highlights the inadequacy of single-gene analysis for diagnostics and surveillance, and underscores the necessity of whole-genome sequencing to accurately track the virus epidemiology and evolution.
