RESUMO
Ionic liquids (ILs) are interesting chemical compounds that have a wide range of industrial and scientific applications. They have extraordinary properties, such as the tunability of many of their physical properties and, accordingly, their activities; and the ease of synthesis methods. Hence, they became important building blocks in catalysis, extraction, electrochemistry, analytics, biotechnology, etc. This study determined antifungal activities of various imidazolium-based ionic liquids against yeast Saccharomyces cerevisiae via minimum inhibitory concentration (MIC) estimation method. Increasing the length of the alkyl group attached to the imidazolium cation, enhanced the antifungal activity of the ILs, as well as their ability of the disruption of the cell membrane integrity. FTIR studies performed on the S. cerevisiae cells treated with the ILs revealed alterations in the biochemical composition of these cells. Interestingly, the alterations in fatty acid content occurred in parallel with the increase in the activity of the molecules upon the increase in the length of the attached alkyl group. This trend was confirmed by statistical analysis and machine learning methodology. The classification of antifungal activities based on FTIR spectra of S. cerevisiae cells yielded a prediction accuracy of 83%, indicating the pharmacy and medicine industries could benefit from machine learning methodology. Furthermore, synthesized ionic compounds exhibit significant potential for pharmaceutical and medical applications.
Assuntos
Antifúngicos , Membrana Celular , Imidazóis , Líquidos Iônicos , Testes de Sensibilidade Microbiana , Saccharomyces cerevisiae , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/química , Líquidos Iônicos/farmacologia , Líquidos Iônicos/química , Imidazóis/farmacologia , Imidazóis/química , Antifúngicos/farmacologia , Antifúngicos/química , Membrana Celular/efeitos dos fármacos , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Fungal infections can be serious or life threatening in severe cases, and the need to discover and find novel antifungal agents persists. Chalcones are plant-derived aromatic compounds that have been appealing synthons for pharmaceutical industry as they have good anticancer, antibacterial, antifungal and anti-inflammatory properties. Although there are few structure-activity relationship studies on chalcones, studies that link the structural features of these compounds to their mode of action are scant. Thus, in this study, we aim to clarify the relationship between chalcone derivatives and their cellular target within the yeast cell Saccharomyces cerevisiae. We observed that some chalcone compounds lead to disruption of cell membrane and cause ion leakage out of the cell. Moreover, chalcones alter the biochemical composition of yeast cells detectable by FTIR spectroscopy and bind to the DNA as shown by our titration experiments based on UV-Vis absorbance spectroscopy. Thus, their interaction with the DNA may be the major impact of these compounds on yeast cells.
Assuntos
Chalcona , Chalconas , Chalcona/farmacologia , Chalconas/farmacologia , Chalconas/química , Saccharomyces cerevisiae , Antifúngicos/farmacologia , Antifúngicos/química , Membrana Celular , Relação Estrutura-Atividade , DNARESUMO
Chalcones have a variety of cellular protective and regulatory functions that may have therapeutic potential in many diseases. In addition, they are considered to affect key metabolic processes in pathogens. Nevertheless, our current knowledge of the action of these compounds against fungal cell is scarce. Therefore, in this study, various substituted chalcone Schiff bases were investigated to reveal their cellular targets within the yeasts Saccharomyces cerevisiae and Candida albicans. First, their antifungal activities were determined via minimum inhibitory concentration method. Surprisingly, parent chalcone Schiff bases showed little or no antifungal activity, while the nitro-substituted derivatives were found to be highly active against yeast cells. Next, we set out to determine the cellular target of active compounds and tested the involvement of the cell wall and cell membrane in this process. Our conductivity assay confirmed that the yeast cell membrane was compromised, and that ion leakage occurred upon treatment with nitro-substituted chalcone Schiff bases. Therefore, the cell membrane came to the fore as a possible target for the active chalcone derivatives. We also showed that exogenous ergosterol added to the growth medium reduced the inhibitory effect of chalcones. Our findings open up new possibilities for the design of future antimicrobial agents based on this appealing backbone structure.
Assuntos
Chalcona , Chalconas , Candida albicans , Chalcona/farmacologia , Saccharomyces cerevisiae , Chalconas/farmacologia , Chalconas/química , Bases de Schiff/farmacologia , Bases de Schiff/química , Antifúngicos/farmacologia , Antifúngicos/química , Testes de Sensibilidade Microbiana , Membrana CelularRESUMO
Chalcones, valuable precursors for flavonoids, have important antibacterial and antifungal activities against bacteria, pathogens, harmful fungi and even antibiotic-resistant microorganisms that cause food spoilage and infectious diseases. It is widely known that chalcones target various vital metabolic pathways of the bacterial cells, but little is known about their action on the cell membrane integrity. In the present study, we studied the antibacterial activity of 12 different substituted chalcones in a comparative way and revealed that the phenolic chalcones are superior to other substituted derivatives against both Gram-negative and Gram-positive bacteria. We also demonstrate that the cell membrane is the first barrier that the chalcone molecules face for their action, and that phenolic chalcones increase ionic cell membrane permeability to a greater extent than the other substituted members. Especially, ion leakage can be detected at lower concentrations than the minimum inhibitory levels against Gram-positive bacteria. Phenolic chalcones are superior to other substituted derivatives in their antibacterial action and cause leakage of ions from Gram-positive bacteria even in concentrations lower than the inhibitory levels. Ion leakage from Gram-positive bacterial cytoplasm is prior to the membrane deformation and cell death. Thus, we propose that ion leakage contribute to the greater activity of phenolic chalcones in comparison to non-phenolic ones, on Gram-positive bacteria. Even though, disruption of metabolic pathways may be the principal mode of action of chalcones; in accord with our observations, we propose that the ion leakage precedes other inhibitory effects and contribute to the antibacterial action of phenolic chalcones.
Assuntos
Chalcona , Chalconas , Morte Celular , Chalconas/farmacologia , Bactérias Gram-Positivas , ÍonsRESUMO
The correct separation of chromosomes during mitosis is necessary to prevent genetic instability and aneuploidy, which are responsible for cancer and other diseases, and it depends on proper centrosome duplication. In a recent study, we found that Smy2 can suppress the essential role of Mps2 in the insertion of yeast centrosome into the nuclear membrane by interacting with Eap1, Scp160, and Asc1 and designated this network as SESA (Smy2, Eap1, Scp160, Asc1). Detailed analysis showed that the SESA network is part of a mechanism which regulates translation of POM34 mRNA. Thus, SESA is a system that suppresses spindle pole body duplication defects by repressing the translation of POM34 mRNA. In this study, we performed a genome-wide screening in order to identify new members of the SESA network and confirmed Dhh1 as a putative member. Dhh1 is a cytoplasmic DEAD-box helicase known to regulate translation. Therefore, we hypothesized that Dhh1 is responsible for the highly selective inhibition of POM34 mRNA by SESA.
Assuntos
RNA Helicases DEAD-box/genética , RNA Helicases DEAD-box/metabolismo , Complexo de Proteínas Formadoras de Poros Nucleares/biossíntese , Mapas de Interação de Proteínas , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Proteínas de Saccharomyces cerevisiae/biossíntese , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/genética , Testes GenéticosRESUMO
OBJECTIVES: Imidazole and benzimidazole derivatives have recently attracted attention as remarkable materials due to their advantages in chemistry, pharmacology, and biomaterials. This article focuses on dental composites with azole functional groups incorporated to affect their physicochemical and mechanical properties and antibacterial activity. METHODS: Dental composites were fabricated by embedding the functionalized imidazole and benzimidazole nanoparticles into a Bis-GMA/TEGDMA matrix to form the imidazole and benzimidazole dental composites series (I and B). The material was produced through hand blending of the monomer (50:50, wt%), filler (0-30, wt%), and initiator combination (CQ/EDMAB:0.8:1.6, wt%), and LED light-curing unit for 60 s. RESULTS: Using various characterization techniques, I and B series were validated. The dental composites' approximate solubility and sorption significances were evaluated by conducting experiments on specific dental composite formulations. Fenton reaction test was performed to determine the chemical stability of the dental composites. The mechanical properties of the dental composites were investigated. Finally, by testing cell growth in the presence of composites, their antibacterial activities were determined. CONCLUSIONS: In this study, it was observed that the mechanical, physiochemical, and antibacterial properties of the functional azole-containing nanoparticles were positively improved by adding them to the structure of dental composites. These experimental results paved the way for the synthesized materials to be used in industrial applications. CLINICAL SIGNIFICANCE: Since the chemical, mechanical, and antimicrobial properties of dental composites containing 10% imidazole and benzimidazole functional nanoparticles are far superior, they constitute an excellent alternative for preventing dental caries and long-term use of dental composites.
Assuntos
Anti-Infecciosos , Cárie Dentária , Nanopartículas , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Azóis , Benzimidazóis , Bis-Fenol A-Glicidil Metacrilato/química , Bis-Fenol A-Glicidil Metacrilato/farmacologia , Resinas Compostas/química , Resinas Compostas/farmacologia , Humanos , Imidazóis , Teste de Materiais , Metacrilatos/química , Nanopartículas/química , Polietilenoglicóis/química , Ácidos Polimetacrílicos/químicaRESUMO
Terpenoids, one of the major components of essential oils, are known to exert potent antifungal activity against yeast Saccharomyces cerevisiae. They have been the subject of a considerable number of investigations that uncovered extensive pharmacological properties, including antifungal and antibacterial effects. However, their mechanism of action remains elusive. In order to use terpenoids as the antimicrobial and antifungal agents in food preservation in a rational way, a good knowledge of their mode of action is required. We hypothesized that the cellular membrane is the main target site for the antifungal agents, and that structural properties of these agents are key to penetrate and act upon phospholipid bilayers. In this study, we thus aimed to study the effect of terpenoids on the cell membrane integrity, with the focus on both their structural properties, such as the presence of aromatic ring or hydroxyl group; and their hydrophobicity, as a consequence of these structural features. We first uncovered the antifungal properties of phenolic terpenoids thymol, carvacrol and eugenol, cyclic terpenes limonene, carveol, and α-pinene, in addition to the closely related compounds of different chemical structures. We then examined the cell membrane deterioration upon the addition of these reagents. Our results demonstrate that the presence of a phenolic -OH moiety is crucial, and hydrophobicity gained by the aromatic ring structure contributes to the ability of penetration and damaging yeast plasma membrane to achieve high antifungal activity.
Assuntos
Antifúngicos/química , Antifúngicos/farmacologia , Membrana Celular/efeitos dos fármacos , Terpenos/química , Terpenos/farmacologia , Membrana Celular/metabolismo , Permeabilidade da Membrana Celular/efeitos dos fármacos , Interações Hidrofóbicas e Hidrofílicas , Testes de Sensibilidade Microbiana , Estrutura Molecular , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Relação Estrutura-AtividadeRESUMO
Correct separation of chromosomes during mitosis is essential for preventing genetic instability and aneuploidy. Such separation is dependent on correct duplication of the nuclear-associated microtubular organizing center, i.e., spindle pole body (SPB), in fungi. MonoPolar Spindle 2 (MPS2) is an essential gene, encoding a membrane protein required for the insertion of SPB into the nuclear envelope. We recently reported that the SESA complex, which is composed of Smy2, Eap1, Scp160, and Asc1, suppresses the essential role of MPS2 (Sezen et al. 2009, Genes & Development 23:1559-1570), i.e., in SESA-active cells Mps2 becomes nonessential. We also proposed that the SESA network facilitates this insertion by altering the membrane lipid composition (Sezen 2015, FEMS Yeast Research 15:fov089). In addition, we are interested in the antifungal properties of essential oils and previously reported that membrane integrity of yeast cells is impaired upon exposure to turpentine, thyme, oregano, and orange peel essential oils (Konuk and Ergüden 2017, BioCell 41:13-18). Due to our continuing interest in the SESA system and the mechanisms by which essential oils affect yeast cells, we aimed to investigate the effects of essential oils on yeast cell membranes. Herein, we show that mps2∆ 2µm-SMY2 and mps2∆ pom34∆ cells, in which the SESA complex is active and SPB duplication is defective, are more prone to membrane damage upon treatment with essential oils.