Epigenetic regulation plays a critical role in controlling fungal secondary metabolism. Here, we report the pleiotropic effects of the epigenetic regulator HdaA (histone deacetylase) on secondary metabolite production and the associated biosynthetic gene clusters (BGCs) expression in the plant endophytic fungus Penicillium chrysogenum Fes1701. Deletion of the hdaA gene in strain Fes1701 induced a significant change of the secondary metabolite profile with the emergence of the bioactive indole alkaloid meleagrin. Simultaneously, more meleagrin/roquefortine-related compounds and less chrysogine were synthesized in the ΔhdaA strain. Transcriptional analysis of relevant gene clusters in ΔhdaA and wild strains indicated that disruption of hdaA had different effects on the expression levels of two BGCs: the meleagrin/roquefortine BGC was upregulated, while the chrysogine BGC was downregulated. Interestingly, transcriptional analysis demonstrated that different functional genes in the same BGC had different responses to the disruption of hdaA. Thereinto, the roqO gene, which encodes a key catalyzing enzyme in meleagrin biosynthesis, showed the highest upregulation in the ΔhdaA strain (84.8-fold). To our knowledge, this is the first report of the upregulation of HdaA inactivation on meleagrin/roquefortine alkaloid production in the endophytic fungus P. chrysogenum. Our results suggest that genetic manipulation based on the epigenetic regulator HdaA is an important strategy for regulating the productions of secondary metabolites and expanding bioactive natural product resources in endophytic fungi.

Owing to the well-established application of prebiotics in human food products, there is a growing interest in their potential as dietary supplements for gut microbiota composition and improvement of the digestive health of dogs. However, targeted studies with dogs as research subjects are still limited. In the present study, an in vitro simulated gut microbiota fermentation system using canine feces from a healthy Border Collie breed was used to investigate the prebiotic effects of five different oligosaccharides and compare their regulatory effects on the gut microbiota structure and the resultant metabolites. Due to the addition of oligosaccharides, the fermented samples had lower pH and higher bacterial proliferation. The oligosaccharide-fermentation selectively boosted Lactobacillus spp., Streptococcus spp., Enterococcus spp., Bacteroides spp., and hindered Escherichia-Shigella spp., Paeniclostridium, spp., and Bacteroides spp. Each oligosaccharide showed distinct characteristics and preferences for regulating gut microbiota structure and abundance. Furthermore, the addition of oligosaccharides increased the production of short-chain fatty acids, particularly butyric acid. This study provides a preliminary basis for the rapid and rational selection of prebiotic oligosaccharides as canine dietary supplements and further explores the function of oligosaccharides and their combinations in canine health.

The quality control of drug products during manufacturing processes is important, particularly the presence of different polymorphic forms in active pharmaceutical ingredients (APIs) during production, which could affect the performance of the formulated products. The objective of this study was to investigate the phase transformation of fexofenadine hydrochloride (FXD) and its influence on the quality and performance of the drug. Water addition was key controlling factor for the polymorphic conversion from Form I to Form II (hydrate) during the wet granulation process of FXD. Water-induced phase transformation of FXD was studied and quantified with XRD and thermal analysis. When FXD was mixed with water, it rapidly converted to Form II, while the conversion is retarded when FXD is formulated with excipients. In addition, the conversion was totally inhibited when the water content was <15% w/w. The relationship between phase transformation and water content was studied at the small scale, and it was also applicable for the scale-up during wet granulation. The effect of phase transition on the FXD tablet performance was investigated by evaluating granule characterization and dissolution behavior. It was shown that, during the transition, the dissolved FXD acted as a binder to improve the properties of granules, such as density and flowability. However, if the water was over added, it can lead to the incomplete release of the FXD during dissolution. In order to balance the quality attributes and the dissolution of granules, the phase transition of FXD and the water amount added should be controlled during wet granulation.

A ternary amorphous solid dispersion (ASD) system consisting of drug/polymer/surfactant is receiving increased attention to improve the oral bioavailability of poorly water-soluble drugs. The effect of polymers has been extensively studied, while the impact of surfactants has not yet to be studied to the same extent. Challenging questions to be answered are whether the surfactants should be added with the drug or separately and the resulting differences between the two operating processes. By adjusting the liquid feeding zone for Span 20 in the hot-melt twin screw extruder equipment, we investigated the effect of Span 20 on the properties of the polyvinylpyrrolidone/vinyl acetate (PVPVA)-based ASD formulations of ritonavir. We found that with the delayed feeding positions of Span 20 in the twin screw extruder, the ability of the ternary ASDs to maintain the supersaturation of the milled extrudates was observed to be significantly enhanced. Furthermore, adding surfactant after a thorough mixing of polymer and drug could decrease the molecular mobility of ternary ASD formulations. In addition, the effects of Span 20 on the complex viscosity and structure of PVPVA were also investigated. The delayed addition of Span 20 could improve the complex viscosity of PVPVA, thus leading to the drug precipitation inhibition. In conclusion, the delayed addition of Span 20 in the twin screw extruder and prolonging the mixing time of the drug and polymer may be critical to the maintenance of supersaturation.

This study selected three approved folate sources—folic acid (FA), L-5-methyltetrahydrofolate (MTFA), and calcium 5-methyltetrahydrofolate (CMTFA)—to explore their interaction mechanisms with soy protein isolate (SPI) through spectrofluorometric analysis and molecular docking simulations. We investigated how these interactions influence the structural and physicochemical stability of folates and SPI. Three folates spontaneously bound to SPI, forming complexes, resulting in a decrease of approximately 30 kJ·mol⁻¹ in Gibbs free energy and an association constant (K_a) of 10⁵ L·mol⁻¹. The thermodynamic parameters and molecular docking study revealed the unique binding mechanisms of FA and MTFA with SPI. FA’s planar pteridine ring and conjugated double bonds facilitate hydrophobic interactions, whereas MTFA’s reduced ring structure and additional polar groups strengthen hydrogen bonding. Although the formation of SPI–folate complexes did not result in substantial alterations to the SPI structure, their binding has the potential to enhance both the physical and thermal stability of the protein by stabilizing its conformation. Notably, compared with free FA, the FA-SPI complexes significantly enhanced FA’s stability, exhibiting 71.1 ± 1.2% stability under light conditions after 9 days and 63.2 ± 2.6% stability in the dark after 60 days. In contrast, no similar effect was observed for MTFA. This discrepancy can be ascribed to the distinct degradation pathways of the Fa and MTFA molecules. This study offers both theoretical and experimental insights into the development of folate-loaded delivery systems utilizing SPI as a matrix.