Toxic Amanita species are a major cause of fatal mushroom poisoning worldwide. Their morphological similarities to the edible species of mushrooms increase the risk of accidental ingestion, highlighting the need for rapid and accurate identification. We aimed to develop a multiplex detection system using isothermal recombinase polymerase amplification (RPA) and CRISPR/Cas12b technology for rapid and specific detection of multiple toxic Amanita species. Species-specific RPA primers and single guide RNAs (sgRNAs) were designed and optimized for six toxic Amanita species (A. exitialis, A. pallidorosea, A. subjunquillea, A. rimosa, A. molliuscula, and A. fuligineoides). The detection system combined RPA with Cas12b-mediated fluorescence readout to evaluate the specificity and sensitivity, and the results were visualized using lateral flow strips and a microfluidic disc chip for parallel detection. The selected primer–sgRNA combinations showed high specificity values without cross-reactivity and a low detection limit of 10 pg of genomic DNA per reaction. The lateral flow strips enabled intuitive and reliable detection. Furthermore, the microfluidic disc chip enabled multiplex detection for both single-species and four-species mixed samples. We successfully established a rapid, sensitive, and specific multiplex detection system for toxic Amanita species using purified genomic DNA from authenticated specimens, providing a proof-of-concept analytical strategy for informing future development of instrument–light species identification tools in mushroom poisoning cases. We also summarize the evaluations required for further optimization of the current design prior to practical application.
Paula Marynella Alves Pereira Lima, André Carlos Pereira de Matos, Gabriela da Silva Guimarães, Laryssa Aparecida Sales Barbosa, Thaise Gonçalves Araújo
Triple-negative breast cancer (TNBC) is a subset of mammary tumors with heterogeneous biology, lacking estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 (HER2). Patients generally exhibit an aggressive course, less favorable disease-specific outcomes, and a higher propensity for early recurrence, with chemotherapy remaining the primary systemic treatment eligible for all patients. In this sense, efforts are urgent to identify new effective compounds, aiming to increase disease-free and overall survival. Copper complexes have emerged as promising in oncology by regulating redox balance and modulating hallmarks such as proliferation, apoptosis, epithelial-mesenchymal transition, and angiogenesis. Furthermore, complexes containing copper tend to have low adverse effects. In TNBC, copper compounds may represent at least one component of a multi-action management strategy, being potentially effective against these tumors. The focus of this review is to provide information on TNBC biology and the role of copper in its signaling pathways, addressing the current copper complexes that have been explored to improve treatment.
Menghui Li, Yihuan Dong, Wen Ren, Weixia Dong, Shaoping Ji
SIRT2, an NAD + -dependent deacetylase, is increasingly recognized as an important regulator of cellular metabolism and aging. By deacetylating diverse substrates, SIRT2 influences glucose metabolism, lipid metabolism, mitochondrial function, cytoskeletal dynamics, oxidative stress, and inflammatory responses, thereby contributing to cellular metabolic homeostasis and aging. Emerging evidence suggests that age-related alterations in NAD + metabolism may affect SIRT2 activity and disrupt these processes, potentially promoting metabolic dysfunction, mitochondrial impairment, cellular senescence, and chronic inflammation. In addition, the effects of SIRT2 on aging appear to be highly context- and tissue-dependent, and its precise contribution to organismal longevity remains incompletely understood. In this review, we primarily focus on the role of SIRT2 in aging and longevity through the regulation of energy metabolism, with particular emphasis on the underlying molecular mechanisms and its interactions with metabolic and stress-responsive pathways. Understanding these mechanisms and regulations may provide new insights into SIRT2 biology and potential strategies for maintaining metabolic homeostasis and promoting healthy aging, even applications in clinic practice.
