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The highly efficient peroxidase‐like activity of platinum nanozymes (3–20 nm size) is exploited within the complex cellular environment to catalyze the oxidation of the DAB substrate, producing an electron‐dense signal around the nanozyme surface, upon osmium staining. It is proved that such nanozyme amplification can achieve a catalytic signal enhancement up to 10‐fold, enabling the quick detection of the Pt particles (even of 3 nm size) by transmission electron microscopy (TEM) also at low magnification and across wide fields of view in the intricate intracellular milieu. The developed procedure is ideally suited to overcome standard amplification strategies currently used in TEM analysis, such as gold or silver enhancements. Furthermore, the wide versatility of the Pt‐nanozyme probes in TEM imaging is demonstrated in immuno‐EM and protein trafficking studies, showing their potential to track the subcellular localization of target biomolecules at both low and high magnifications. These results suggest that the use of nanozymes might represent a paradigm shift in the conventional amplification systems currently employed in electron microscopy for cellular analyses, offering enhanced imaging capabilities. [ABSTRACT FROM AUTHOR]

The cytotoxic action of anticancer drugs can be potentiated by inhibiting DNA repair mechanisms. RAD51 is a crucial protein for genomic stability due to its critical role in the homologous recombination (HR) pathway. BRCA2 assists RAD51 fibrillation and defibrillation in the cytoplasm and nucleus and assists its nuclear transport. BRC4 is a peptide derived from the fourth BRC repeat of BRCA2, and it lacks the nuclear localization sequence. Here, we used BRC4 to (i) reverse RAD51 fibrillation; (ii) avoid the nuclear transport of RAD51; and (iii) inhibit HR and enhance the efficacy of chemotherapeutic treatments. Specifically, using static and dynamic light scattering, transmission electron microscopy, and microscale thermophoresis, we show that BRC4 eroded RAD51 fibrils from their termini through a "domino" mechanism and yielded monomeric RAD51 with a cumulative nanomolar affinity. Using cellular assays (BxPC-3, pancreatic cancer), we show that a myristoylated BRC4 (designed for a more efficient cell entry) abolished the formation of nuclear RAD51 foci. The present study provides a molecular description of RAD51 defibrillation, an essential step in BRCA2-mediated homologous recombination and DNA repair. [ABSTRACT FROM AUTHOR]

The ultrastructure of mature spermatozoa from three leech species, Haemadipsa zeylanica ( Moquin-Tandon, 1826 ) (Haemadipsidae), Theromyzon tessulatum ( Müller, 1774 ) and Placobdella costata (Fr. Müller, 1846 ) (Glossiphoniidae), was investigated by means of transmission electron microscopy. While the three species showed the general features of hirudinid spermatozoa, considerable variation was found in nearly all characters examined: the anterior acrosome varied in length and in shape; the nuclei also varied in size and show a very complicated shape which is maintained for the whole length in the two glossiphonids, while in the haemadipsid, it showed two different morphological regions. The single mitochondrion, which is surrounded by an electron-dense sheath, is straight in H and T. tessulatum, but twisted in P. costata. The flagellum in the three studied species is of the prominent central sheath type. The basal body is absent in the three species and the central apparatus of the flagellum was observed penetrating to the mitochondrion. By comparing the present data with those from “oligochaeta”, Branchiobdellida and Acanthobdellida, we found that only the presence of an anterior acrosome characterizes the true leeches and at present should be regarded as an autapomorphic character of Euhirudinea. [ABSTRACT FROM AUTHOR]

A method for the aqueous synthesis of stable and biocompatible citrate-coated palladium nanoparticles (PdNPs) in the size range comparable to natural enzymes (4–8 nm) has been developed. The toxicological profile of PdNPs was assessed by different assays on several cell lines demonstrating their safety in vitro also at high particle concentrations. To elucidate their cellular fate upon uptake, the localization of PdNPs was analyzed by Transmission Electron Microscopy (TEM). Moreover, crucial information about their intracellular stability and oxidation state was obtained by Sputtering-Enabled Intracellular X-ray Photoelectron Spectroscopy (SEI-XPS). TEM/XPS results showed significant stability of PdNPs in the cellular environment, an important feature for their biocompatibility and potential for biomedical applications. On the catalytic side, these PdNPs exhibited strong and broad antioxidant activities, being able to mimic the three main antioxidant cellular enzymes, i.e., peroxidase, catalase, and superoxide dismutase. Remarkably, using an experimental model of a human oxidative stress-related disease, we demonstrated the effectiveness of PdNPs as antioxidant nanozymes within the cellular environment, showing that they are able to completely re-establish the physiological Reactive Oxygen Species (ROS) levels in highly compromised intracellular redox conditions. [ABSTRACT FROM AUTHOR]