Myocardial ischemia-reperfusion injury (MIRI) poses a major clinical challenge. This study aimed to elucidate whether TRPM7 regulates cardiomyocyte pyroptosis during MIRI and its underlying mechanisms. We established both in vivo (I/R-injured mice) and in vitro (H/R-injured HL-1 cardiomyocytes) models and employed multiple techniques including gene knockdown/overexpression, specific inhibitors (KN‑93 for CaMKII inhibition), protein‑protein interaction assays (Co‑IP, GST pull‑down), and m5C modification analysis (MeRIP, dual‑luciferase reporter assay with site‑directed mutagenesis). The results demonstrated that TRPM7 knockdown significantly improved cardiac function, reduced infarct area and myocardial fibrosis, and suppressed cell death in mice. Mechanistically, TRPM7‑mediated Ca2+ influx activated CaMKII, which bound to and upregulated DDX3X. Concurrently, I/R‑ or H/R‑induced m5C modification on DDX3X mRNA recruited the m5C reader protein ALYREF, thereby stabilizing DDX3X expression. The elevated DDX3X subsequently activated the NLRP3 inflammasome, ultimately driving cardiomyocyte pyroptosis. Importantly, in vivo knockdown of DDX3X recapitulated the cardioprotective effects observed with TRPM7 inhibition. In conclusion, our findings reveal a critical role of a TRPM7‑initiated CaMKII/DDX3X/NLRP3 signaling axis in regulating pyroptosis during MIRI, providing a novel potential therapeutic target for this condition.
DNA methylation changes are recognized as a key hallmark of biological aging. We analyzed the products of active DNA demethylation pathways in DNA isolated from leukocytes and the urine of healthy volunteers aged between 1.5 and 71 years. A statistically significant inverse correlation between age and the analyzed modifications in urine was found for 5-hydroxymethyluracil, 5-hydroxymethylcytosine, 5-(hydroxymethyl)-2′-deoxycytidine, 5-methyl-2′-deoxycytidine, 5-carboxycytosine and 5-formylcytosine. The highest correlation coefficient was observed for 5-hydroxymethyluracil (r = − 0.66). Interestingly, statistically significant inverse correlations were observed for 5-(hydroxymethyl)-2′-deoxycytidine and 5-methyl-2′-deoxycytidine in DNA. Changes in epigenetic modifications with age are more pronounced in urine than in cellular DNA. However, DNA modifications are limited to a single tissue (leukocytes), whereas modifications detected in urine reflect processes occurring at the level of the entire organism.
Parent-offspring conflict is a fundamental concept in evolutionary biology that explains the discrepancy between parents’ and offspring’s optimal strategies for allocating parental investments. Two alternative evolutionary strategies were proposed to explain the timing of termination of the post-fledging dependence period (PFDP). The age-of-independence hypothesis (H1) predicts that offspring will receive parental care for as long as possible, as determined by parents. The early-dispersal hypothesis (H2) states that young attain independence as soon as possible once reaching ontogenetic maturity, as determined by offspring. We radio-tracked 91 Tengmalm’s owl (Aegolius funereus) fledglings from 31 nests over six years in Czechia and Finland until independence. The mean, minimum and maximum recorded PFDP durations were longer in Czechia [49.4 ± 6.4 days (mean ± SD; range: 34–61 days)] than in Finland [38.3 ± 6.5 days (28–54 days)]. In both areas, the PFDP duration was relatively short (44.5 ± 4.9 vs. 36.6 ± 4.8 days, respectively) during the rich, but much longer (56.7 ± 3.4 vs. 50.5 ± 2.5 days, respectively) during the poor season of food abundance. The PFDP was shorter across sites with delayed hatching dates, probably reflecting the higher quality of early-nesting parents and their ability to invest more in offspring, as well as the need for all parents to recover from parental effort before winter onsets. The longer PFDPs during food shortage, and in Czechia compared to Finland, as well as the shorter PFDP duration with delayed offspring hatching dates, supported the age-of-independence hypothesis (H1), but were inconsistent with the early-dispersal hypothesis (H2). Therefore, we suggest that parents primarily decide on the length and termination of PFDP. We suggest that maintaining prolonged PFDP parental care is important for Tengmalm’s owl offspring’s prospects, enabling the development of flight skills that directly impact hunting success after reaching independence, the critical period for their future survival. Finally, our results demonstrate, for the first time, that differing environmental conditions across distant study sites (Central versus North Europe) are crucial in determining PFDP duration, an important life-history trait of altricial birds.
This study analyzed the content and composition of phenolic compounds, flavonoids, coumarins, pigments, and neuroactive secondary metabolites in fly agaric (Amanita muscaria) fruiting bodies collected in August and September 2022 and 2023 from two sites: under pine trees and under birch trees in the Wielkopolska region. A total of 15 phenolic compounds, as well as groups of flavonoids and other bioactive metabolites, including ergosterol, were determined. The study aimed to evaluate how environmental factors—such as the type of symbiotic tree, seasonal differences, and interannual variation in temperature and rainfall—affect the biosynthesis of these metabolites. The results showed significantly higher concentrations of most analyzed compounds in fruiting bodies collected under pine trees, particularly in 2023, which had higher temperatures and lower rainfall. Seasonal variation was evident: higher concentrations of phenols and flavonoids were found in August, while ergosterol and secondary pigments increased in September, suggesting shifts in metabolic activity and fruiting body maturation. Coumarins were also more abundant in pine-associated samples, indicating enhanced production of protective metabolites under stress conditions. Toxic alkaloids such as ibotenic acid and muscimol were present at higher levels in pine fruiting bodies, though their concentrations varied seasonally. Statistical analysis confirmed that both the year and habitat had significant effects on metabolite content, with clear differences in profiles between sites and years. The study results confirm that the species of symbiotic tree and weather conditions (year and month of collection) significantly influence the biosynthesis of phenolic compounds, flavonoids, and coumarins in A. muscaria fruiting bodies. Mycorrhiza with pine and the warmer weather of 2023 favored increased synthesis of protective compounds. Seasonal monitoring (August vs. September) provides valuable information on the metabolic dynamics of fungi and may be useful in the search for bioactive compounds. Furthermore, total phenolic content (TPC) and antioxidant activity (ABTS·+) can serve as indicators of environmental stress and the antioxidant potential of fly agaric fruiting bodies.
