An unprecedented effort to tackle the ongoing COVID-19 pandemic has characterized the activity of the global scientific community over the last two years. Hundreds of published studies have focused on the comprehension of the immune response to the virus and on the definition of the functional role of SARS-CoV-2 proteins. Proteins containing zinc fingers, both belonging to SARS-CoV-2 or to the host, play critical roles in COVID-19 participating in antiviral defenses and regulation of viral life cycle. Differentially expressed zinc finger proteins and their distinct activities could thus be important in determining the severity of the disease and represent important targets for drug development. Therefore, we here review the mechanisms of action of host and viral zinc finger proteins in COVID-19 as a contribution to the comprehension of the disease and also highlight strategies for therapeutic developments.

Ethanol exposure during pregnancy alters the mammalian target of rapamycin (mTOR) signaling pathway in the fetal brain. Hence, in adult rats exposed to ethanol during the neonatal period, we investigated the influence of rapamycin, an mTOR Complex 1 (mTORC1) inhibitor, on deficits in spatial memory and reversal learning in the Barnes maze task, as well as the ethanol-induced rewarding effects (1.0 or 1.5 g/kg) using the conditioning place preference (CPP) paradigm. Rapamycin (3 and 10 mg/kg) was given before intragastric ethanol (5 g/kg/day) administration at postnatal day (PND)4–9 (an equivalent to the third trimester of human pregnancy). Spatial memory/reversal learning and rewarding ethanol effect were evaluated in adult (PND60–70) rats. Additionally, the impact of rapamycin pre-treatment on the expression of the GluN2B subunit of NMDA receptor in the brain was assessed in adult rats. Our results show that neonatal ethanol exposure induced deficits in spatial memory and reversal learning in adulthood, but the reversal learning outcome may have been due to spatial learning impairments rather than cognitive flexibility impairments. Furthermore, in adulthood the ethanol treated rats were also more sensitive to the rewarding effect of ethanol than the control group. Rapamycin prevented the neonatal effect of ethanol and normalized the GluN2B down-regulation in the hippocampus and the prefrontal cortex, as well as normalized this subunit’s up-regulation in the striatum of adult rats. Our results suggest that rapamycin and related drugs may hold promise as a preventive therapy for fetal alcohol spectrum disorders.

Brain microvascular endothelial cells experience hypoxic conditions in several neurodegenerative disease processes and the underlying mechanisms still need to be explored. Current imaging modalities and biochemical assays require many specific markers that should be detected to identify the hypoxic response, especially at a level of single cells. This study presents a single-cell molecular imaging approach utilizing Fourier-Transform Infrared and Raman spectroscopy. Those methods enable the simultaneous detection of proteins, lipids, and nucleic acids encoded in their unique vibrational fingerprints. By establishing ratiometric estimators, we measured upregulated lipid metabolism, structural changes of proteins and asses DNA:RNA ratio at the single-cell level induced by oxygen depletion. Moreover, this approach allows for analyzing changes within specific cellular compartments, including nuclei, providing a comprehensive understanding of how hypoxia affects cellular functions and metabolism. Our findings pave the way for future investigations into the cellular adaptations to hypoxia in brain endothelial cells, potentially revealing novel therapeutic targets for neurodegenerative diseases.

Classic infrared (IR) microscopy is limited by the diffraction limit, which obscures subcellular heterogeneity, and by the complex overlap of vibrational bands, which complicates precise molecular assignment. This study presents a comprehensive "IR map of the cell" that provides a standardized framework for label-free chemical identification of subcellular compartments across various human cell lines. By integrating Fourier transform infrared (FTIR) spectroscopy with submicron-resolution optical photothermal infrared (OPTIR) microscopy (∼0.3 μm), the cellular landscape was mapped with improved spatial specificity beyond that achievable by conventional FTIR imaging. Advanced chemometric tools were employed to segment the nucleus, cytoplasm, and regions rich in lipids and glycogen, each characterized by a definitive "IR barcode". Furthermore, in silico modeling validated spectral assignments by simulating cellular fingerprints from reference biocompounds, while detailed spectroscopic characterization of subcellular compartments defined marker bands for proteins, lipids, carbohydrates, and nucleic acids. The modeling supported the interpretation that experimental spectra can be approximated as a linear combination of biomolecular classes, helping to constrain spectral band overlap and refine the proposed "IR barcode" for cellular identification. The developed IR map provides a robust, standardized foundation for the label-free interpretation of cellular chemistry. This study demonstrates the utility of IR microscopy as a powerful diagnostic and analytical tool for monitoring metabolic shifts and cellular status at the micrometric level.

Mass spectrometry imaging (MSI) is a method for surface analysis that was introduced at the end of the twentieth century, and since then a huge development of this technique can be observed. It is widely used in biological, biochemical, and medical research. MSI technique combines two elements. The first is the ability to obtain the mass spectrum of a given point on the surface, which provides information about the molecules present in this particular point. The second element is the design of the ion source, which enables movement of the examined surface in the x- and y-axis. The most widespread MS techniques for imaging are secondary ion mass spectrometry, matrix-assisted laser desorption/ionization, and desorption electrospray ionization. Each of them allows for the analysis of slightly different types of molecules. The chapter discusses their advantages and disadvantages in the context of surface imaging as well as exemplary applications.

The blood–brain barrier (BBB) is a coordinated multicellular interface in which intercellular communication shapes responses to injury and metabolic stress. However, the contributions of individual cell types to collective stress adaptation remain poorly understood. Here, we examined molecular and metabolic responses of 2D BBB models comprising endothelial cells, astrocytes, and pericytes exposed to hypoxia or sublethal HgCl₂ toxicity. An integrated platform combining gene and protein expression profiling, label-free vibrational spectroscopic imaging, and machine-learning-based analysis was used to assess stress-induced changes in cell-cycle regulation and metabolism across monoculture and co-culture configurations. Hypoxia broadly suppressed G1/S and G2/M regulators, inducing cell-cycle arrest with limited cell death, consistent with an adaptive response. HgCl₂ caused heterogeneous, model-dependent changes in viability and proliferative capacity. Spectroscopic phenotyping showed that co-cultured cells acquired molecular profiles characteristic of pericytes, indicating a dominant pericyte-driven influence on the cellular community. Fluorescence imaging further revealed intercellular mitochondrial transfer at cell–cell contact sites. These findings identify pericytes as key contributors to stress resistance and BBB cell survival. The developed spectroscopic–molecular platform provides a label-free framework for monitoring cellular communication and stress responses in multicellular BBB models.