Journal of Molecular Medicine, Journal Year: 2024, Volume and Issue: unknown
Published: Sept. 25, 2024
Language: Английский
FEBS Open Bio, Journal Year: 2025, Volume and Issue: unknown
Published: April 3, 2025
Macroautophagy/autophagy is a crucial cellular process for degrading and recycling damaged proteins organelles, playing significant role in diseases such as cancer neurodegeneration. Evaluating autophagy flux, which tracks autophagosome formation, maturation, degradation, essential understanding disease mechanisms. Current fluorescence‐based methods are resource‐intensive, requiring advanced equipment expertise, limiting their use clinical laboratories. Here, we introduce non‐fluorescent immunohistochemistry (IHC) method using MAP1LC3/LC3 SQSTM1 core markers flux assessment. LC3 levels reflect whereas degradation decrease the number of its puncta indicate active (i.e., lysosomal turnover). We optimized chromogenic detection diaminobenzidine (DAB) staining developed scoring system based on percentage stained cells. This accessible, cost‐effective enables reliable quantification standard light microscope, bridging gap between experimental research diagnostics. Our protocol allows accurate evaluation fixed tissues, offering practical applications biomedical pathology
Language: Английский
Citations
0
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown
Published: June 30, 2024
Abstract Macroautophagy/autophagy, a crucial cellular process, is typically measured using fluorescence-based techniques, which can be costly, complex, and impractical for clinical settings. In this paper, we introduce novel, cost-effective, non-fluorescent immunohistochemistry (IHC) method evaluating autophagy flux. This technique, based on antigen-antibody reactions chromogenic detection, provides clear, quantifiable results under standard light microscopy, eliminating the need expensive equipment specialized reagents. Our simplifies technical requirements, making it accessible to routine laboratories research settings with limited resources. By comparing our approach traditional fluorescence methods, demonstrate its superior effectiveness, cost-efficiency, applicability patient samples. innovative technique has potential significantly advance improve diagnostics, offering practical robust tool studying mechanisms in diseases such as cancer neurodegenerative disorders. IHC represents significant step forward flux, more reliable, promise of enhancing understanding treatment autophagy-related diseases.
Language: Английский
Citations
2
iScience, Journal Year: 2024, Volume and Issue: 27(8), P. 110447 - 110447
Published: July 4, 2024
Early childhood caries (ECC) is a multifactorial disease with microbiome playing significant role in progression. Understanding changes at the level ECC required to develop diagnostic and preventive strategies. In our study, we combined data from small independent cohorts compare composition using unified pipeline applied batch correction avoid pitfalls of effects. Our meta-analysis identified common biomarker species between different studies. We best machine learning method for classification versus caries-free samples compared performance this leave-one-dataset-out approach. random forest model was found be generalizable when used combination other While results highlight potential microbial involved classification, also mentioned limitations that can serve as guide future researchers design use appropriate tools such analyses.
Language: Английский
Citations
1
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown
Published: Aug. 10, 2024
Abstract Autophagy is a critical cellular process involved in the degradation and recycling of cytoplasmic components, playing dual role cancer by either promoting cell survival or facilitating death. In glioblastoma (GB), autophagy has been implicated resistance to chemotherapeutic agent Temozolomide (TMZ). This study presents novel method accurately measure flux TMZ-resistant cells, combining advanced imaging techniques with biochemical assays. By quantifying key markers such as LC3-II SQSTM1, our approach provides detailed insights into dynamic processes autophagosome formation clearance under therapeutic stress. not only advances understanding GB chemoresistance but also significant implications for development autophagy-targeted therapies. The ability monitor manipulate real-time offers promising avenue monitoring understnading TMZ improving patient outcomes treatment.
Language: Английский
Citations
1
Methods in molecular biology, Journal Year: 2024, Volume and Issue: unknown
Published: Jan. 1, 2024
Language: Английский
Citations
1
Biochemistry and Biophysics Reports, Journal Year: 2024, Volume and Issue: 39, P. 101752 - 101752
Published: June 1, 2024
Language: Английский
Citations
0
Journal of Molecular Medicine, Journal Year: 2024, Volume and Issue: unknown
Published: Sept. 25, 2024
Language: Английский
Citations
0