Brian T.

UB Center of Infectious Diseases, University at Buffalo, Buffalo, NY 14214, USA School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, Buffalo, NY 14214, USA

Bacterial immune systems exhibit remarkable diversity and modularity, as a consequence of the continuous selective pressures imposed by phage preda...

Multiple primate species, including humans, evolved brains that are exceptionally large relative to their body sizes. These large brains coevolved ...

Background/Objectives: Botanical supplements are increasingly investigated for their potential to address women’s health concerns. Compounds that modulate progesterone receptor (PR) signaling may help manage gynecologic disorders such as endometriosis, uterine hyperplasia, and preterm birth. Because PR ligands often cross-react with the glucocorticoid receptor (GR), this study examined two botanical compounds, paeoniflorin from Paeonia lactiflora (white peony root) and isoorientin from Vitex agnus-castus (chasteberry), that were identified as modulators of GR or PR signaling. Methods: Luciferase reporter assays were performed in OVCAR5, Ishikawa PR-B, and T47D A1-2 cells to evaluate GR and PR signaling. GR target gene expression was measured by qPCR. A receptor binding assay and computational docking were used to assess interaction with GR. Adipogenesis was evaluated in 3T3-L1 cells using Oil Red O staining and FABP4 protein expression by Western blot. Results: Paeoniflorin and isoorientin inhibited dexamethasone-放induced GR signaling in OVCAR5 and Ishikawa PR-B cells. In T47D A1-2 cells, a variant of T47D engineered to express GR, both compounds blocked luciferase induction stimulated by progesterone; this effect was not observed in the parental line that expresses PR but lacks GR. In OVCAR5 cells, paeoniflorin or isoorientin combined with dexamethasone downregulated GILZ and DUSP1/MKP1 mRNA. Isoorientin directly bound GR, and computational analysis supported potential binding poses. Both compounds also reduced lipid accumulation during 3T3-L1 adipocyte differentiation and decreased FABP4 expression, consistent with GR antagonist activity and reduced adipogenesis. Conclusions: These findings identify paeoniflorin and isoorientin as botanical modulators that suppress GR signaling and limit GR-dependent adipogenic responses across multiple cell-based models under controlled in vitro conditions.

Background and rationale In our ongoing phase II observational pilot trial, the compounded formulation of low-dose rapamycin significantly reduced fatigue-related clinical symptoms in ME/CFS subjects. Although the underlying molecular mechanism remains unclear, exploring metabolic pathways involving circulating blood-borne factors is warranted. Recent studies suggest that increased levels of purines may exacerbate oxidative stress in ME/CFS patients. It is not known if rapamycin modulates purine biosynthesis and improves disease symptoms. Methods and results To address, we performed a comprehensive LCMS-based quantification of purine biosynthetic intermediates in plasma from responder cohort of ME/CFS participants, both at baseline (BSL) and after 90 days of rapamycin therapy (T3). Notably, differential regulation was observed in the enzymatic conversion of inosine monophosphate (IMP) to xanthosine-5-monophosphate (XMP) and hypoxanthine (HPX) between BSL and T3 samples. Flow cytometry assays on PBMCs confirmed that rapamycin reduces IMP dehydrogenase activity, thereby limiting the conversion of IMP to XMP. Further analyses, including mitochondrial oxidative stress assessments, Seahorse OCR following purine supplementation, and flow cytometry indicate that altered purine levels can impair mitochondrial energy metabolism, and may contribute to inflammatory processes in microglia. Conclusion Collectively, these findings highlight the therapeutic potential of rapamycin to enhance energy metabolism in patients with ME/CFS. Major limitations There is no placebo group, and molecular results are somewhat biased to responders. Trial registration CLINICALTRIALS.GOV, NCT06257420. Registered 11 December 2023, https://clinicaltrials.gov/study/NCT06257420.