Chronic sleep deprivation (SD) disrupts central nervous system homeostasis, promoting behavioral alterations and glial remodeling within limbic structures. Although simvastatin exhibits pleiotropic effects, its influence on astrocytic and microglial responses during prolonged sleep loss remains poorly understood. This study investigated the effects of SD and simvastatin on behavior, systemic inflammatory mediators, and glial reactivity in the dorsal hippocampus (HPC) and amygdala (AMY). Sixty male Wistar rats (~75 days old; n = 15/group) were assigned to Control (CG), Sleep Deprived (SD), Simvastatin 4.2 mg/kg (SIM4.2), or Simvastatin 20 mg/kg (SIM20) groups. Animals underwent the modified multiple platform method (18 h/day) for 45 consecutive days, with daily oral simvastatin administration. Social behavior was assessed using the resident-intruder test, circulating cytokines by ELISA, and glial responses by GFAP and IBA-1 immunohistochemistry, including microglial morphological classification (Types I-III). SD increased aggressive/reactive behavior and astrocytic and microglial reactivity in the dorsal HPC, accompanied by a shift from homeostatic (Type I) toward reactive (Types II and III) microglial phenotypes. Simvastatin attenuated these glial alterations, restoring a more homeostatic microglial profile and increasing non-social exploratory behavior. In contrast, glial alterations in the AMY were comparatively limited, indicating region-specific susceptibility to SD. Peripheral cytokine analysis revealed selective immunomodulatory effects, including reduced TGF-β and increased IL-8 in SIM20, whereas TNF-α and IL-6 remained unchanged. Overall, simvastatin modulated behavioral, systemic inflammatory, and glial responses to SD, supporting its pleiotropic effects and highlighting region-specific glial remodeling as a potential target for mitigating the consequences of chronic sleep loss.

Pleurotus ostreatus is a well-known culinary-medicinal mushroom for its rich nutritional composition and diverse biological activities. The present study aimed to comparatively evaluate 115 wild isolates and five commercial strains cultivated under controlled bottle-culture conditions in terms of β-glucan, ergothioneine, and lovastatin contents, as well as selected biological activities and nutritional parameters. Isolates PO85, PO71, and PO36 exhibited the highest β-glucan [61.526 g/100 g dry weight (dw)], lovastatin (1.509 mg/g dw), and ergothioneine (10.732 mg/g dw) contents, respectively. In biological activity assays, the isolate PO85 promoted the growth of Lactobacillus strains effectively, lovastatin-rich extract of isolate PO116 pronounced antifungal activity against Candida albicans, and ergothioneine-rich extract of isolate PO164 exhibited the highest antioxidant capacity in the ABTS radical scavenging assay. In terms of nutritional analysis, isolate PO71 was rich in dry matter, nitrogen, and protein, whereas isolate PO85 had high moisture, ash, and reducing sugar levels but lower fat content. These findings highlight the significant intraspecific variability of P. ostreatus and suggest that some isolates may offer potential for future functional food applications due to their metabolite profiles and biological activities.

This study demonstrates the considerable antagonistic activity of Aspergillus terreus MAK153 against a pathogenic Fusarium oxysporum AUMC3191 isolate. In dual culture assay, soil-isolated Aspergillus terreus Thom effectively hindered F. oxysporum AUMC3191 growth and impressively blocked zearalenone (ZEN) synthesis. Scanning electron microscopy demonstrated significant damage to F. oxysporum hyphae, such as abnormalities, shrinkage, bending, and deformity. Molecular identification through rDNA ITS sequence analysis confirmed the isolate as Aspergillus terreus, registered in GeneBank (Accession No. OR480103). GC/MS profiling of A. terreus MAK153 identified several bioactive secondary metabolites, notably estragole, caryophyllene, 2,4-di-tert-butylphenol, 1-docosene, 1-docosanol, olean-12-ene-3,28-diol, and simvastatin. Statistical analysis revealed a very strong positive correlation (r = 0.99) between chromatographic peak area% and peak area values. The influence of nutrient sources on simvastatin production was evaluated using four culture media. FTIR and HPLC analyses confirmed simvastatin biosynthesis, while HPLC revealed significant medium-dependent variation in yield (p < 0.001). Camellia sinensis Tea broth (CTB) elicited the highest yield, dominating extracellular filtrate accumulation (145.93 ± 1.13 mg/mL). Molecular docking simulations assessed simvastatin's binding affinity to HMG-CoA reductase and 3-ketosteroid reductase, yielding binding energies of -8.0 and -7.8 kcal/mol, respectively. The interactions involved multiple hydrophobic bonds and hydrogen bonds, suggesting simvastatin functions as a multi-target inhibitor capable of disrupting ergosterol biosynthesis at two distinct sites. Notably, this study represents the first computational exploration of simvastatin's interaction with 3-ketosteroid reductase, with implications for estrogen signalling modulation and potential therapeutic applications. KEY POINTS: • Aspergillus terreus MAK153 inhibits F. oxysporum growth and zearalenone production • Aspergillus terreus induces significant hyphal damage in F. oxysporum using SEM • Simvastatin-HMG-CoA and 3-ketosteroid interaction could modulate estrogen-related pathways potentially.

Hyperlipidemia is an important cardiovascular risk factor, particularly in individuals with a strong family history of hypercholesterolemia and premature cardiovascular disease. Red yeast rice (RYR) contains monacolin K, a naturally occurring compound structurally similar to lovastatin, and has demonstrated lipid-lowering effects. This case describes a 50-year-old man with hyperlipidemia and clinical features raising suspicion for an inherited lipid disorder who declined statin therapy. Over 12 months, the patient implemented dietary and lifestyle modifications and used RYR and other nutritional supplements. Total cholesterol decreased from 243 to 187 mg/dL, low-density lipoprotein cholesterol from 166 to 114 mg/dL, and triglycerides from 123 to 76 mg/dL, while high-density lipoprotein cholesterol increased from 55 to 58 mg/dL. Fasting glucose decreased from 100 to 92 mg/dL, and HbA1c was 5.4% when measured at follow-up. No adverse effects from the nutritional supplements were reported during follow-up. This case describes improvement in lipid parameters during a multimodal nutritional, lifestyle, and supplemental intervention and highlights the need for further research to evaluate these approaches in individuals with hyperlipidemia.

This study examined the effects of purified lovastatin (LV) obtained from Monascus fermentation on ruminal fermentation, methane production, and microbial communities during a 48-h in vitro incubation. A fattening-sheep diet containing concentrate and roughage at 60:40 was treated with 0, 50, 100, or 150 mg LV/kg dry matter (DM). Samples from the control and 100 mg/kg DM treatments were selected for bacterial and archaeal 16S rRNA amplicon sequencing. Compared with the control, supplementation with 100 mg LV/kg DM significantly decreased methane production (mL/g DM) and methane content (%) by 22.1% and 12.7%, respectively, while increasing in vitro DM degradation by 4.9%. Cumulative gas production and total VFA concentration did not differ significantly among treatments. The molar proportion of propionate was higher at 150 mg/kg DM, while MCP concentration was higher at 100 mg/kg DM. Rikenellaceae RC9 gut group, Eubacterium, Succinivibrionaceae UCG-001, and Anaerovibrio were enriched in the LV treatment, whereas Methanococcus and Methanosphaera were enriched in the control. These results demonstrate the potential of lovastatin as a natural inhibitor of ruminal methanogenesis, with 100 mg/kg DM showing the greatest efficacy among the doses tested under the present in vitro conditions.

Pharmaceutical pollutants that contain lactone or lactam rings in their structure are frequently detected in wastewater. Enzymes with lactonase activity, synthesized by the microorganisms in aerobic activated sludge, are expected to be able to participate in the hydrolysis of these pharmaceutical pollutants, but the actual activity of many lactonases towards such compounds remains poorly characterized. In this work, a systematic molecular docking study of 12 enzymes and 39 pharmaceutical pollutants (β-lactam antibiotics, macrolides, coumarins, statins) was undertaken. The binding affinity, active-site occupancy, total enzyme surface occupancy and geometric characteristics of the molecules (the distance from the catalytic nucleophile to the carbonyl carbon of the lactone ring, and the attack angle formed between the nucleophilic atom of the enzyme and the C-N/C-O bond of the substrate) were evaluated in silico for a set of hydrolases displaying lactonase activity (AidC, AaL, hexahistidine-tagged organophosphate hydrolase (His6-OPH), etc.) and metallo-β-lactamases (MβLs) (NDM-1, MIM-1, SAM-1). Lactamases exhibited favorable intermolecular interaction geometry for the majority of β-lactams. No enzyme was found that satisfied the geometric criteria corresponding to the biodegradation of macrolides. With coumarins, only Paraoxonase PON2 provided an optimal geometry of biocatalysis; however, this enzyme is absent from activated sludge but may be of interest for in vivo applications. For several statins widely used in practice, combinations with enzymes having virtually ideal geometry for biocatalysis were identified: AidC/rosuvastatin lactone, AaL/lovastatin, His6-OPH/mevastatin, and PON2/simvastatin. For the first time, several pharmaceutical lactones were identified in silico and confirmed in vitro as inhibitors of MβL and His6-OPH. Competitive inhibition of NDM-1 by lactone-containing signaling molecules of Gram-negative bacteria present in activated sludge was revealed for the first time both in silico and in vitro. The obtained results' may be taken into account when implementing monitoring of biological stages at water treatment stations and improve the efficiency of water purification. The revealed characteristics of MβL inhibition by macrolides may be taken into account as one of possible explanations for the effective application of β-lactams in combinations with macrolides known from multiple paratactical investigations.

Monacolin K (MK) in red yeast rice (RYR) is structurally identical to pharmaceutical lovastatin in its lactone form, making their origins difficult to distinguish. This study examined the δ[13]C value of hydrolysis-derived 2-methylbutyric acid (2-MBA) as a fragment-specific screening marker using liquid chromatography-isotope ratio mass spectrometry (LC-IRMS). Within-day repeatability was 0.19‱ for RYR, interlaboratory standard deviations were 0.18-0.19‱, and a two-component expanded uncertainty estimate was ±0.58‱ (k = 2). Hydrolysis-derived 2-MBA ranged from -32.73‱ to -26.68‱ in 45 authentic RYR samples and from -10.17‱ to -6.87‱ in 23 pharmaceutical lovastatin batches. Controlled blends showed a linear relationship between δ[13]C and blend ratio (R[2] = 0.9997). A one-sided tolerance limit at 95% confidence and 99% population coverage gave an investigational screening threshold of -26.47‱ for the sampled Chinese authentic RYR population. Three of 20 commercial products exceeded this threshold and were classified as isotopically inconsistent with that population. Overall, this study developed and analytically evaluated a fragment-specific LC-IRMS approach for screening source-related isotope differences between RYR-derived MK and pharmaceutical lovastatin under the investigated conditions, providing a potential analytical tool for RYR authenticity assessment.

Using transformation-associated recombination (TAR) technology, a lovastatin biosynthetic gene cluster was assembled in Saccharomyces cerevisiae BY4741, in which pathway-related enzyme genes were controlled by strong fungal promoters and terminators. Using Aspergillus niger SH-2 as the starting strain, a copy of Monacolin J (MJ) and a copy of lovastatin expression cassette were integrated into its genome, MJ heterologous expression strains and lovastatin heterologous expression strains were successively constructed. To facilitate the synthesis of MJ and lovastatin, ATP-dependent citrate lyase (ACL) and acetyl-CoA carboxylase (ACC) were overexpressed in the MJ- and lovastatin-engineered strains. For fermentation optimization, various carbon sources were tested to determine the best one. Furthermore, the roles of glutamic acid, methionine, and linoleic acid in the production of MJ and lovastatin were investigated. Finally, in a 500 mL shake flask with 100 mL of liquid medium, using hydrolyzed starch as the carbon source and adding 10 mM glutamate, a MJ titer of 1606 mg/L and a lovastatin of 1345 mg/L were obtained, and after overexpressing ACL and ACC enzymes in the MJ strain, a MJ titer of 1786 mg/L were obtained. During fed-batch fermentation in a 2-L bioreactor, the lovastatin yield reached 2915 mg/L.

Sex differences in immunity shape cancer risk, autoimmunity, and responses to immunotherapy, yet the sex-chromosome genes that regulate antitumor T cell function remain incompletely defined. Here, we identify the Y chromosome-encoded KDM5D histone demethylase as a male-specific suppressor of CD8+ T cell antitumor immunity. In murine colorectal cancer (CRC) models, male CD8+ T cells displayed reduced cytokine production, proliferation, cytotoxicity, TCRβ abundance, and proximal TCR signaling relative to female CD8+ T cells. CRISPR-RNP-mediated KDM5D depletion in male CD8+ T cells enhanced effector function, increased TCRβ expression, augmented TCR signaling, and improved tumor control after adoptive transfer. Transcriptomic and functional analyses further linked KDM5D to cholesterol biosynthesis and exhaustion-associated programs, with KDM5D depletion reducing SREBP2/XBP1-associated cholesterol and exhaustion signatures. Correspondingly, human CRC single-cell analyses supported the clinical relevance of this axis, showing enrichment of exhausted and cholesterol-associated CD8+ T cell states in male tumors. Pharmacologic inhibition of cholesterol biosynthesis with lovastatin partially attenuated select exhaustion-associated markers in male CD8+ T cells and delayed tumor growth in vivo. Together, these findings define KDM5D as a sex chromosome-encoded regulator of male CD8+ T cell dysfunction and point to cholesterol-exhaustion programs as a potential therapeutic vulnerability in male CRC.

Liver fibrosis in metabolic dysfunction associated steatohepatitis (MASH) lacks effective therapies, in part because therapeutic agents poorly penetrate the capillarized sinusoidal endothelium. As fibrosis progresses, liver sinusoidal endothelial cells (LSECs) become defenestrated and restrict trans-sinusoidal exchange, thereby limiting drug access to hepatic stellate cells (HSCs) within the perisinusoidal space. Here, we developed a lipoprotein mimetic liposomal nanocarrier, SV@SRB-Lip, to exploit scavenger receptor class B type 1 (SR-B1)-associated transendothelial transport across capillarized LSECs and enhance downstream delivery toward HSC-associated fibrotic regions. SV@SRB-Lip was engineered as a simvastatin loaded liposomal platform incorporating an ApoA-I-mimetic SRB peptide, an MMP-responsive linker, and a vitamin A modified lipid moiety. SV@SRB-Lip promoted transport across the LSEC monolayer in vitro, and this effect was reduced by SR-B1 silencing, supporting the involvement of SR-B1-associated endothelial transport. In MASH mice, SV@SRB-Lip enhanced hepatic delivery and distributed toward HSC associated fibrotic regions. The MMP-responsive design increased the relative accessibility of vitamin A within the fibrotic microenvironment, supporting downstream HSC associated uptake. As a result, SV@SRB-Lip increased hepatic simvastatin accumulation, suppressed HSC activation and collagen deposition, and improved MASH associated fibrosis. These findings suggest that SR-B1-associated endothelial transport can be harnessed to overcome the capillarized sinusoidal barrier and improve antifibrotic nanomedicine delivery in MASH.

Citrus species differ widely in flavonoid chemistry, yet grapefruit has come to represent the genus in citrus-statin interactions. To test whether this translates into species-dependent interaction profiles, six flavonoid-profiled citrus peel extracts (Citrus limon, C. depressa Hayata, C. maxima, C. × paradisi, C. × sinensis, and C. reticulata) were compared in a standardized acute rat model spanning drug disposition, CYP3A and transporter systems, hepatic redox status, and in silico analysis. The naringin-dominated C. × paradisi produced the most coordinated disruption of CYP3A and transporter pathways, raising plasma simvastatin acid at 1 h by 506% versus the simvastatin-only group, together with the least favorable redox profile. The polymethoxyflavone-rich C. depressa Hayata perturbed disposition mildly yet raised the hepatic GSH/GSSG ratio by 227%. Hepatic active-metabolite accumulation was therefore uncoupled from redox outcome, indicating that disposition and redox response are separable axes and that grapefruit cannot represent the citrus genus.

The Extended Clearance Classification System (ECCS) predicts predominant systemic clearance mechanisms of small-molecule drugs based on molecular weight, ionization, and permeability. Highly permeable acids and zwitterions with a molecular weight exceeding 400 Da (ECCS class 1B) are expected to rely on transporter-mediated hepatic uptake. For this class, human hepatic clearance is frequently underpredicted when extrapolated from in vitro metabolic stability, complicating prospective pharmacokinetic predictions. Here, we systematically evaluated the performance of metabolism-based intrinsic hepatic clearance predictions using curated clinical intravenous pharmacokinetic data for 16 ECCS class 1B drugs. Although approximately 70% of compounds were predicted within 3-fold of observed values, clinically meaningful underprediction persisted for a subset (atorvastatin, bosentan, pitavastatin, lovastatin acid, and telmisartan). To prospectively identify compounds at risk of >3-fold underprediction, we compared 4 approaches: (1) rat in vitro-in vivo correlation (IVIVC), (2) nonhuman primate (NHP) IVIVC, (3) IVIVC in chimeric mice with humanized livers (PXB-mice), and (4) measurement of the fraction of active uptake in suspended human hepatocytes. Rat IVIVC reliably identified compounds scaling adequately in humans but showed lower discriminatory power to identify compounds with >3-fold underprediction. In contrast, a human hepatocyte active uptake fraction >50% robustly identified the compounds with human hepatic clearance underprediction. NHP and PXB-mouse IVIVC provided lower discriminatory power. For compounds flagged to be at risk of >3-fold underprediction by the hepatic uptake assay, incorporation of measured hepatic uptake clearance into scaling quantitatively improved human clearance predictions, achieving accuracy comparable to single-species NHP allometric scaling. SIGNIFICANCE STATEMENT: Accurate clearance prediction for Extended Clearance Classification System class 1B drugs is complicated by transporter-mediated hepatic uptake, which can cause substantial underprediction when scaling from metabolic stability. This work introduces a practical decision framework combining rat in vitro-in vivo correlation and human hepatocyte uptake screening to identify compounds at risk of >3-fold clearance underprediction. For the identified compounds, uptake-informed scaling improves human pharmacokinetic predictions, reduces uncertainty during lead optimization, and lowers reliance on nonhuman primates in drug discovery.