Search Results (57)

G-quadruplex (DG4) is folded in guanine-rich DNA sequences and regulates DNA replication and transcription. Although bioinformatics analyses have predicted the presence of DG4 in maize, its biological functions remain largely unexplored. In this study, we treated maize seedlings with 0, 100, 200, and 300 μM TMPyP4, a DG4-stabilizing ligand, and observed that TMPyP4 inhibits radicle growth by increasing reactive oxygen species (ROS) levels and inducing DNA fragmentation in radicle cells. Transcriptomic RNA-seq revealed that TMPyP4 modulated the expression of 1614 genes in maize radicle cells, which were predominantly associated with redox reactions, membrane components, and secondary metabolic pathways. BG4-ChIP-seq analysis demonstrated that DG4 structures are evenly distributed across the ten chromosomes of the maize genome, occupying 22,449 loci and showing significant enrichment for specific DG4-binding motifs. Integrative analysis of RNA-seq data and BG4-ChIP-seq identified 944 differentially expressed genes, which were significantly enriched in pathways related to redox reactions and secondary metabolism. Collectively, these findings suggest that DG4-stabilizing ligands regulate maize radicle growth by modulating ROS homeostasis, providing critical insights into the functional roles of DG4s in maize. Full article

Antimicrobial photodynamic therapy is a method that utilizes photodynamic inactivation of microorganisms exposed to a photosensitizer irradiated by a specific wavelength, followed by the formation of reactive oxygen species and subsequent oxidative stress. In contrast to antibiotics, which are generally efficient against specific microorganisms, photodynamic inactivation exhibits efficacy against a wide range of bacteria, representing a promising and non-invasive alternative to treating infections caused by pathogens of different origins. This study compares the antibacterial efficacy of five different photosensitizers, including TMPyP, Protoporphyrin IX, PdTPPS₄, Methylene Blue, and ZnPCS₂, against eight representatives of various pathogens, including Gram-negative bacteria Escherichia coli, Pseudomonas aeruginosa, Gram-positive bacteria Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium, MRSA and Bacillus subtilis. An optimal irradiation protocol was developed based on growth curve measurements involving double irradiation. To induce the photodynamic effect, we utilized LED emitters with wavelengths of 414 nm and 660 nm, chosen to align with the photophysical properties of the photosensitizers. Additionally, the research included assessments of the radiation’s phototoxicity and the photosensitizers’ dark toxicity against specific microorganisms. The optical properties of the photosensitizers were analyzed using absorption spectrophotometry. The effectiveness of photodynamic inactivation was assessed by determining the minimum inhibitory and bactericidal concentrations. This study aimed to identify the most suitable photosensitizer for clinical application, considering the toxicity of the photosensitizer, the radiant exposure, and its efficacy in photodynamic inactivation. Full article

This study focuses on the rational design and synthesis of new pyrrolidinone derivatives aimed at inhibiting the ACE2 receptor, a key entry point for SARS-CoV-2. Building on structural insights from known inhibitors like MLN-4760, five compounds featuring a pyrrolidinone core, with some incorporating indole rings, were synthesized and characterized. Their inhibitory activity against ACE2 was evaluated in vitro, with compound 4 showing the most promising results, further confirmed through cytotoxicity assays and molecular docking studies. Docking analysis revealed favorable binding interactions within the ACE2 active site, supporting the potential of compound 4 as a therapeutic agent. These findings provide a foundation for future in vivo studies and highlight the potential development of novel therapeutics targeting ACE2-related diseases, including COVID-19. Full article

Endothelial cell (EC) barrier integrity is tightly regulated by the activity of the non-muscle myosin light chain kinase (nmMLCK) under diverse pathological inflammatory conditions (pneumonia, sepsis) and exposure to mechanical stress. Inflammatory stimuli, including lipopolysaccharide (LPS), cytokines, and damage-associated molecular patterns (DAMPs), increase EC permeability through nmMLCK-dependent EC paracellular gap formation. However, the exact mechanisms by which nmMLCK regulates vascular barrier dysfunction in acute lung injury (ALI) remain incompletely understood. We hypothesized that inflammation-induced ROS results in the peroxynitrite-mediated nitration of nmMLCK that contributes to EC barrier disruption. Human lung EC exposure to either the peroxynitrite donor, SIN-1, or to LPS, triggered significant nmMLCK nitration, which was abolished by the oxidant scavenger, MnTMPyP. Mass spectrometry of SIN-1-treated nmMLCK identified multiple nitrated tyrosines. Nitration of Y1410 proved a critical PTM as site-directed substitution with alanine (Y1410A) abolished both SIN-1- and LPS-induced nmMLCK nitration. nmMLCK nitration disrupts wild-type nmMLCK interaction with Kindlin-2, a cytoskeletal regulator of vascular barrier stability, whereas EC transfected with the Y1410A nmMLCK mutant exhibited preserved Kindlin-2 binding, reflected by alterations in trans-EC electrical resistance (TEER). Consistent with these observations, LPS-challenged murine lungs displayed enhanced nmMLCK nitration and diminished nmMLCK-Kindlin-2 association. Functionally, SIN-1 markedly impaired EC barrier integrity (TEER), which was not observed in ECs expressing the Y1410A mutant. Together, these findings suggest that nmMLCK nitration at Y1410 is a critical molecular mechanism contributing to vascular leakage, highlighting this modification as a potential therapeutic target to reduce inflammation-induced vascular permeability. Given nmMLCK’s established role in barrier regulation, we hypothesized that LPS-induced peroxynitrite formation may promote the nitration of nmMLCK tyrosine residues: a PTM that potentially contribute to nmMLCK’s regulation of EC barrier integrity. Full article

Porphyrins play an extremely important role in both photodynamic (PDT) and sonodynamic therapy (SDT). These techniques, which have a lot in common, are largely based on the interaction between the sensitizer and light or ultrasounds (US), respectively, resulting in the formation of reactive oxygen species (ROS) that have the ability to destroy target cells. SDT requires the use of an appropriate frequency of US waves that are able to excite the chemical compound used. In this study, five porphyrin complexes were used: free-base meso-tetra(N-methyl-pyridinium-4-yl)porphyrin (TMPyP) and its transition metal complexes containing zinc(II), palladium(II), copper(II), and chloride-iron(II). The sonodynamic activity of these compounds was studied in vitro. The obtained results confirm the significant relationship between the chemical structure of the macrocycle and its stability and ability to generate ROS. The highest efficiency in ROS generation and high stability were demonstrated by non-metalated compound and its complex with zinc(II), while complex with copper(II), although less stable, were equally effective in terms of ROS production. Antibacterial activity tests showed the unique properties of the tested compounds, including a reduction in the number of both planktonic and biofilm antibiotic-resistant microorganisms above 5 log, which is rare among sonosensitizers. Full article

The photodynamic inactivation (PDI) of bacteria is a promising alternative to antibiotics in boar semen extenders. It was recently established using the illumination of semen samples containing 2 µM of the photosensitizer 5,10,15,20-tetrakis(N-methyl-4-pyridyl)-21H,23H-porphine (TMPyP) with white LED light. High concentrations of TMPyP require strict sample handling in the dark to avoid uncontrolled photodynamic effects caused by ambient light. This study was designed to examine whether lower concentrations of PS could be utilized along with a narrow band blue LED light source, which aligns with TMPyP’s Soret band, thereby minimizing light-induced disruption. A dose-response study with blue LED light exposure of sperm revealed no light toxicity. Importantly, substituting the established white light PDI with blue light illumination and 0.5 µM TMPyP resulted in robust antimicrobial efficiency and sperm compatibility in long-term stored semen samples. This modification led to the confirmation of the hypothesis that a diminished TMPyP concentration in concert with blue LED light facilitates semen handling in normal laboratory light while avoiding unintended light effects. In conclusion, this study plays a pivotal role in augmenting the practicality of the innovative PDI technology by establishing a method that is less susceptible to unanticipated effects of ambient light during sample management. Full article

Singlet oxygen quantum yield (Φ_Δ) is a critical parameter in photodynamic studies, particularly for evaluating photosensitizers’ efficiency in diverse applications such as photodynamic therapy and environmental remediation. Standard photosensitizers, including Rose Bengal, Methylene Blue, and porphyrins, are widely employed as benchmarks for determining Φ_Δ. However, accurate determination of Φ_Δ relies not only on the intrinsic properties of these photosensitizers but also on their experimental conditions, such as concentration. This study investigated the influence of photosensitizer concentration on singlet oxygen quantum yield using several standard photosensitizers. Our findings revealed a significant decrease in Φ_Δ with increasing photosensitizer concentrations across all tested compounds. This decline was attributed to self-quenching effects and molecular aggregation, which reduced the efficiency of energy transfer from the excited triplet state of the photosensitizer to molecular oxygen. The results emphasize the importance of optimizing photosensitizer concentration to ensure reliable Φ_Δ measurements and avoid underestimations. This work underscores the need to consider concentration-dependent effects in future studies to ensure accurate and reproducible outcomes. Full article

Leishmania major is the etiological agent of cutaneous leishmaniasis (CL) in several countries in Asia and Northern Africa. The disease is considered a zoonotic infection where rodents are the reservoirs and phlebotomine sandflies are the vectors. Once inside the human body, the parasite multiplies inside the macrophages of infected patients, but the disease eventually cures spontaneously, leaving scars where the phlebotomine bites occurred. Given the importance of the replicative forms in the parasite’s cell cycle, here, we decided to study the enzyme telomerase, which has the critical role of replenishing the chromosomal telomeric ends during cell replication. To this aim, we first conducted partial purification using Sephacryl-300 HR gel filtration, which allowed us to determine that the telomerase activity eluted as a 600 KDa complex. Second, we characterized an immunopurified L. major telomerase, and to try to explain some of our findings, we performed modeling studies using Alfa fold 3, Pyre2, and Swiss Protein Model. Finally, considering the similarity between the catalytic site of Leishmania and Homo sapiens telomerase, we decided to test typical inhibitors of human telomerase on the purified enzyme and promastigote cell forms, confirming that MST-312 and TMPYP4 efficiently inhibited L. major activity and arrested cell growth in Leishmania promastigotes. Our findings confirm the importance of telomerase activity in L. major’s replicative forms and suggest the possibility of using drugs previously tested on human telomerase to treat CL. Full article

Cancer is a leading cause of death, so continuous efforts into cancer therapy are imperative. In tumor cells, telomerase and oncogene activity are key points for uncontrolled cell growth. Targeting these processes with ligands that inhibit telomerase and/or reduce oncogene expression has been identified as a promising cancer therapy. This study evaluated the selectivity and affinity of the silver^II complex of 5,10,15,20-tetrakis(N-methyl-4-pyridinium)porphyrin (AgTMPyP) to stabilize DNA sequences capable of forming G4 structures mimicking the telomeric and oncogene regions, using spectroscopic, biochemical methods and in vitro assays. The tetracationic silver complex was compared with the free base, H₂TMPyP, and the zinc^II complex, ZnTMPyP. The results obtained from UV-Vis and fluorescence methods pointed to a great affinity and good selectivity of AgTMPyP to G4 structures, especially for the oncogene MYC. In general, an increase in the ability of the studied ligands for ¹O₂ generation when interacting with oncogenic and telomeric G4 sequences was found. The results of the PCR stop assays proved that AgTMPyP has the ability to inhibit Taq polymerase. Additionally, in vitro assays demonstrated that the silver^II complex exhibits low cytotoxicity against HaCaT— an immortalized, non-tumorigenic, skin keratinocytes cell line—and, although nonexclusive, AgTMPyP shows nuclear co-localization. Full article

G-quadruplexes (G4s) are non-canonical nucleic acid secondary structures that have gathered significant interest in medicinal chemistry over the past two decades due to their unique structural features and potential roles in a variety of biological processes and disorders. Traditionally, research efforts have focused on stabilizing G4s, while in recent years, the attention has progressively shifted to G4 destabilization, unveiling new therapeutic perspectives. This review provides an in-depth overview of recent advances in the development of small molecules, starting with the controversial role of TMPyP4. Moreover, we described effective metal complexes in addition to G4-disrupting small molecules as well as good G4 stabilizing ligands that can destabilize G4s in response to external stimuli. Finally, we presented antisense strategies as a promising approach for destabilizing G4s, with a particular focus on 2′-OMe antisense oligonucleotide, peptide nucleic acid, and locked nucleic acid. Overall, this review emphasizes the importance of understanding G4 dynamics as well as ongoing efforts to develop selective G4-unfolding strategies that can modulate their biological function and therapeutic potential. Full article

Porphyrins are frequently employed in photodynamic therapy (PDT), a non-invasive technique primarily utilized to treat subcutaneous cancers, as photosensitizing agents (PAs). The development of a new PA with improved tissue selectivity and efficacy is crucial for expanding the application of PDT for the management of diverse cancers. We investigated the systemic effects of 5,10,15,20-tetrakis(N-methylpyridinium-3-yl)-porphyrin (TMPyP3) using Drosophila melanogaster adult males. We established the oral administration schedule and demonstrated that TMPyP3 was absorbed and stored higher in neuronal than in non-neuronal extracts. Twenty-four hours after oral TMPyP3 photoactivation, the quantity of hydrogen peroxide (H₂O₂) increased, but exclusively in the head extracts. Regardless of photoactivation, TMPyP3 resulted in a reduced concentration of H₂O₂ after 7 days, and this was linked with a decreased capacity to climb, as indicated by negative geotaxis. The findings imply that systemic TMPyP3 therapy may disrupt redox regulation, impairing cellular signaling and behavioral outcomes in the process. To determine the disruptive effect of porphyrins on redox homeostasis, its duration, and the mechanistic variations in retention across various tissues, more research is required. Full article

The G-quadruplex is one of the non-canonical structures formed by nucleic acids, which can be formed by guanine-rich sequences. They became the focus of much research when they were found in several oncogene promoter regions and also in the telomeres. Later on, they were discovered in viruses as well. Various ligands have been developed in order to stabilize DNA G-quadruplexes, which were believed to have an anti-cancer or antiviral effect. We investigated three of these ligands, and whether they can also affect the stability of the G-quadruplex-forming sequences of the RNA genome of SARS-CoV-2. All three investigated oligonucleotides showed the G-quadruplex form. We characterized their stability and measured their thermodynamic parameters using the Förster resonance energy transfer method. The addition of the ligands caused an increase in the unfolding temperature, but this effect was smaller compared to that found earlier in the case of G-quadruplexes of the hepatitis B virus, which has a DNA genome. Full article

Water-soluble metalloporphyrins, depending on the metal center, possess special spectral, coordination, and photochemical features. In nickel(II) porphyrins, the Ni(II) center can occur with low-spin or high-spin electronic configuration. In aqueous solution, the cationic nickel(II) complex (Ni(II)TMPyP⁴⁺, where H₂TMPyP⁴⁺ = 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin), exists in both forms in equilibrium. In this study, an equilibrium system involving the low-spin and high-spin forms of Ni(II)TMPyP⁴⁺ was investigated via application of irradiation, temperature change, and various potential axial ligands. Soret band excitation of this aqueous system, in the absence of additional axial ligands, resulted in a shift in the equilibrium toward the low-spin species due to the removal of axial solvent ligands. The kinetics and the thermodynamics of the processes were also studied via determination of the rate and equilibrium constants, as well as the ΔS, ΔH, and ΔG values. Temperature increase had a similar effect. The equilibrium of the spin isomers was also shifted by decreasing the solvent polarity (using n-propanol) as well as by the addition of a stronger coordinating axial ligand (such as ammonia). Since triethanolamine is an efficient electron donor in Ni(II)TMPyP⁴⁺-based photocatalytic systems, its interaction with this metalloporphyin was also studied. The results promote the development of efficient photocatalytic systems based on this complex. Full article

G-quadruplexes (G4s) are noncanonical nucleic acid structures that play significant roles in regulating various biological processes, including replication, transcription, translation, and recombination. Recent studies have identified G4s in the genomes of several viruses, such as herpes viruses, hepatitis viruses, and human coronaviruses. These structures are implicated in regulating viral transcription, replication, and virion production, influencing viral infectivity and pathogenesis. G4-stabilizing ligands, like TMPyP4, PhenDC3, and BRACO19, show potential antiviral properties by targeting and stabilizing G4 structures, inhibiting essential viral life-cycle processes. This review delves into the existing literature on G4’s involvement in viral regulation, emphasizing specific G4-stabilizing ligands. While progress has been made in understanding how these ligands regulate viruses, further research is needed to elucidate the mechanisms through which G4s impact viral processes. More research is necessary to develop G4-stabilizing ligands as novel antiviral agents. The increasing body of literature underscores the importance of G4s in viral biology and the development of innovative therapeutic strategies against viral infections. Despite some ligands’ known regulatory effects on viruses, a deeper comprehension of the multifaceted impact of G4s on viral processes is essential. This review advocates for intensified research to unravel the intricate relationship between G4s and viral processes, paving the way for novel antiviral treatments. Full article