Nuclear factor of activated T cells 5 (NFAT5) is a transcription factor within the Rel family, primarily recognized for its role in cellular adaptation to osmotic stress, particularly in hypertonic and hyperosmotic environments. Beyond osmotic regulation, NFAT5 responds to diverse stimuli, including cytokines, growth factors, oxidative stress, and microbial signals. This versatility enables NFAT5 to regulate essential cellular processes such as proliferation, survival, migration, and vascular remodelling. In the immune system, NFAT5 modulates the function of monocytes, macrophages, astrocytes, microglia, and T cells, contributing to immune homeostasis and inflammatory responses. Dysregulation of NFAT5 activity is implicated in various pathological conditions, including autoimmune diseases, cancer, and cardiovascular disorders, largely due to its ability to control genes involved in inflammatory and immune pathways under both isotonic and hypertonic conditions. Recent studies have unveiled new regulatory mechanisms, including interactions with non-coding RNAs, offering deeper insights into the functional landscape of NFAT5 and its therapeutic potential. This review delves into the multifaceted roles of NFAT5 in health and disease, emphasizing its emerging importance as a promising therapeutic target.

Differential and even opposing functions of two major antioxidant transcription factors Nrf1 and Nrf2 (encoded by Nfe2l1 and Nfe2l2, respectively) are determined by distinctions in their tempospatial positioning, topological repartitioning, proteolytic processing, and biochemical modification, as well as in their shared evolutionary origin. As a matter of fact, the allelopathic potentials of Nrf1 and Nrf2 (both resembling two entangled ‘Yin-Yang’ quanta that comply with a dialectic law of the unity of opposites) are fulfilled to coordinately control redox physiological homeostasis so as to be maintained within the presetting thresholds. By putative exponential curves of redox stress and intrinsic anti-redox capability, there is inferable to exist a set point at approaching zero with the ‘Golden Mean’ for the healthy survival (i.e., dubbed the ‘zero theory’). A bulk of the hitherto accumulating evidence demonstrates that the set point of redox homeostasis is dictated selectively by multi-hierarchical threshold settings, in which the living fossil-like Nrf1 acts as a robust indispensable determinon, whereas Nrf2 serves as a versatile chameleon-like master regulon, in governing the redox homeodynamic ranges. This is attributable to the facts that Nrf2 has exerted certain ‘double-edged sword’ effects on life process, whereas Nrf1 executes its essential physiobiological functions, along with unique pathophysiological phenotypes, by integrating its ‘three-in-one’ roles elicited as a specific triplet of direct sensor, transducer and effector within multi-hierarchical stress responsive signaling to redox metabolism and target gene reprogramming. Here, we also critically reviewed redox regulation of physio-pathological functions from the eco-evo-devo perspectives, through those coding rules (redox code, stress-coping code, and topogenetic code). The evolving concepts on stress and redox stress were also further revisited by scientific principles of physics and chemistry. Besides, several novel concepts such as oncoprotists, Reverse Central Dogma, and Grand Redox-Unifying Theory’ (GRUT) of life, together with diffusive reactive species (DRS)-based murburn concept integrating all stochastic electron-, proton- and/or moiety-transfer reactive and interactive processes (e.g., PCHEMS), are introduced in this interdisciplinary and synthetic review.

Metabolic dysfunction-associated steatotic liver disease (MASLD), formerly referred to non-alcoholic fatty liver disease (NAFLD), has become a global health concern with a still increasing prevalence. One of the major contributing factors to its pathogenesis is overnutrition. In recent years, a discussion has been started that not only general overnutrition but also specific dietary patterns like the so-called ‘Western diet’ composed of foods rich in saturated fats, cholesterol, and sugar (especially fructose) but low in fiber and polyunsaturated fats, may contribute to the development of MASLD. Evidence from human (intervention) studies regarding the effects of sugar and especially fructose intake is limited and contradictory with respect to the development of MASLD. Still, some scientific liver societies have incorporated a reduction of sugar-sweetened beverages (SSBs) being rich in fructose in their life-style advice for the treatment of MASLD. Being metabolized independently of insulin, fructose has been proposed to be processed more rapidly than glucose, leading to increased lipogenesis and subsequently to hepatic lipid accumulation. Results of more recent experimental studies suggest that an elevated intake of fructose may also affect gut microbiota composition, alter small intestinal morphology and impair intestinal barrier function subsequently leading to an increased translocation of pathogen-associated molecular patterns (PAMPs) into the portal circulation. In this narrative review we summarize recent findings related to the relationship of fructose intake and MASLD, herein focusing on the gut-liver axis and the discrepancy between studies in humans and model organisms.

The human gut microbiota, a diverse community of beneficial normal flora microorganisms, significantly influences physiological function and the immune response. Various microbiota strains have shown promise in supporting clinical treatment of chronic diseases, including cancer, by potentially providing antioxidative and anti-tumorigenic effects in both in vivo and in vitro studies. Breast cancer, which ranks amongst the top five cancer types common worldwide and particularly in Mediterranean countries, has been showing high incidence and prevalence. In breast cancer, microbiota composition, hormonal dynamics, and dietary choices are believed to play significant roles. Hence, the Mediterranean diet, known for its microbiota-friendly features, emerges as a potential protective factor against breast cancer development, highlighting the potential for personalized dietary strategies in cancer prevention. This comprehensive review highlights the emerging mechanisms by which probiotics support our immune system during different physiological activities. It also discusses their potential role, along with nutrition intervention, in improving essential clinical treatment outcomes in breast cancer patients and survivors, suggesting potential supportive strategies that go hand in hand with clinical strategies. Unfortunately, very little research addresses the possible clinical implications of probiotics and dietary habits on breast cancer, despite the promising results, calling for further studies and actions.

The dysfunction of mitochondria, the “energy factories” of cells, not only causes an insufficiency of energy production but also leads to various pathological alterations in cells such as the accumulation of reactive oxygen species, inflammatory responses and mitochondrial DNA damage, all of which were involved in the onset or deterioration of diseases. The presence of mitochondrial dysfunction has been confirmed in many ocular surface diseases such as dry eye, Fuchs corneal endothelial dystrophy and diabetic keratopathy. However, its role in the pathogenesis of ocular surface diseases and underlying molecular mechanisms have not been fully elucidated. Moreover, mitochondrial therapies for ocular surface diseases are currently still under investigation. This mini-review summarizes the pathological features of mitochondrial dysfunction and its mechanisms that have been identified in the pathogenesis of ocular surface diseases, and discusses the potential of mitochondrial therapies in the treatment of these diseases.

Genetically identical cells in a population show cell-to-cell variability because of fluctuation in transcription, epigenetics, post-translational modifications, and stochastic or extrinsically triggered non-genetic alterations. The change in the interaction state of proteins also emerges as an additional layer of cell signaling that influences cell cycle and cell death. However, the interrelation between cell cycle progression and cell death under the influence of spatio-temporal changes in protein-protein interaction is difficult to demonstrate in growing cells. This requires tools for cell cycle phase-resolved visualization of macromolecular interactions in live cells. We describe an approach to visualize the interaction of pro- and anti-death signaling partners, Bax and Bcl-xL, during cell cycle progression and cell death in live cells. Cells were stably expressed with Bax and Bcl-xL with FRET pairs and real-time cell cycle indicator probes. Acceptor photobleaching and Fluorescence lifetime imaging revealed interaction dynamics between Bax and Bcl-xL in isogenic stable cells. While Bcl-xL inhibited cell cycle progression, Bax promoted the cell cycle. The study highlighted an increased Bax-Bcl-xL interaction in the G1 phase compared to the non-G1 phase. Increased interaction is seen under stressed conditions and Bax-activated cells with FLIM-FRET, highlighting the nature of Bax-Bcl-xL interaction during cellular stress. In conclusion, our study explains Bax-Bcl-xL interaction dynamics in real-time and the potential utility of the approach to study macromolecular interactions along with cell cycle and cell death.

An imbalanced production of reactive oxygen species (ROS) is linked to various aspects of cancer development, including cytoskeletal remodelling. However, the relationship between ROS, actin and cellular stiffness remains controversial. Here, we show that oxidative stress increases cortical stiffness in pre-apoptotic colon and pancreatic cancer cells via localized F-actin polymerization in the apical cortex — independent of changes in total F-actin levels. Using atomic force microscopy and flow cytometry, we demonstrate this effect across multiple ROS inducers: the combination of arsenic trioxide and D-enantiomer of vitamin C, hydrogen peroxide, and rotenone. These findings explain previously debated relationships on how ROS influence actin organization, which may affect cellular stiffness. By separating total from cortical actin effects, our study reveals a redox-sensitive mechanism that governs cytoskeletal remodelling and may impair cancer cell migration.

Replication stress (RS) is a major driver of genomic instability and cancer development through impaired DNA replication that can lead to chromo-somal instability (CIN). Although RS is mechanistically linked to CIN, its relationship with cellular proliferation is complex. Depending on the con-text, RS can either promote or suppress cell growth. Existing RS gene expression signatures overlook this complexity, relying on the overex-pression of oncogenes such as MYC, which introduces a proliferation bias. To disentangle genuine RS from confounding cell cycle and prolifer-ation transcriptional profiles, we developed and validated a novel gene expression signature that accurately predicts RS independently of onco-gene activity. This tumorigenic RS signature (TRSS) captures RS-related transcriptional changes across diverse cellular contexts, enabling a more robust and proliferation-independent measure of RS in both experimental and clinical samples. Applying our signature to patient data, we discov-ered a link between RS and the non-homologous end-joining (NHEJ) DNA repair pathway. Specifically, we observed that MSH2 and MSH6 – core components of mismatch repair – are associated with elevated RS and may indicate a shift toward NHEJ-mediated repair under stress condi-tions. Our study provides a refined approach to quantify RS and sheds light on its broader impact on DNA repair network dynamics.

High carbohydrate intake, a characteristic of many Western diets, is a major contributor to age-associated pathologies. Here, we explored the molecular consequences of sugar overload during chronological aging in the yeast Saccharomyces cerevisiae. High levels of glucose and fructose resulted in a decrease of chronological lifespan as well as an increase of cell death, ROS and neutral lipids. Interestingly, these changes were accompanied by significantly altered ceramide profiles. Deletion of either the kinase Tor1, a master regulator of growth and autophagy in response to nutrients, or the vacuole-anchored receptor Vac8, an important player in various autophagy pathways, improved survival and normalized ceramide profiles. This suggests that ceramides might play a role in sugar stress-induced cell death. In line, pharmacological inhibition of sphingolipid synthesis normalized ceramide profiles and improved chronological lifespan, whereas pharmacologically induced ceramide accumulation decreased chronological lifespan. In sum, our findings causally link nutrient signaling and an altered ceramide profile to sugar cytotoxicity in aging yeast, providing a basis for further search of feasible interventions against sugar-induced cell death.

Antibody-drug conjugates (ADCs) offer a strategy for targeted delivery of cytotoxic agents to cancer cells. In this study, we investigated the mechanism of action of datopotamab deruxtecan, an ADC composed of a monoclonal antibody targeting tumor-associated calcium signal transducer 2 (TACSTD2, also known as trophoblast cell-surface antigen-2 (TROP2)) conjugated to the topoisomerase I inhibitor DXd. Datopotamab deruxtecan reduced the viability of human osteosarcoma U2OS cells engineered to express TROP2, but had no effect on their parental counterparts, which only expressed the CALR-GFP biosensor. In TROP2-expressing cells, it triggered the translocation of CALR-GFP from the ER to the cell periphery. Both datopotamab deruxtecan and its DXd payload elicited several features characteristic of immunogenic cell death (ICD), including detectable calreticulin exposure on the cell surface, release of high-mobility group box 1 (HMGB1), and ATP secretion into the culture medium. Importantly, the TROP2-targeted ADC also exerted a bystander antitumor effect on parental U2OS cells (lacking TROP2 expression) co-cultured with TROP2-expressing U2OS cells. These findings demonstrate that datopotamab deruxtecan delivers a cytotoxic payload capable of inducing hallmark features of ICD in vitro.