RESUMO
The management of invasive fungal infections in humans poses significant challenges due to the intricate nature of the treatment, which is both arduous and costly, necessitating routine diagnostic procedures. Consequently, this investigation aimed to formulate a chitosan-based nanoemulsion (CS NEMs) incorporating the antifungal agent undecanoic acid (UDA), characterizing these NEMs and assessing their antifungal efficacy against both filamentous and non-filamentous fungal pathogens. The CS-based UDA NEMs were synthesized by introducing the surfactant Triton X-100 and the stabilizer glycerol. Nanoparticle tracking analysis (NTA) and SEM demonstrated the CS-UDA NEMs with an average size of 145 nm and 164.5 ± 24 nm, respectively. The successful formation of CS-UDA NEMs was verified through FTIR and XRD. CS-UDA NEMs exhibited exceptional inhibition against Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, and Candida albicans with MFC of 500, 500, 250 and 250 µg/mL, respectively. Additionally, CS-UDA NEMs displayed comparatively lower antioxidant activity as determined by DPPH and ABTS radical scavenging assays. Importantly, CS-UDA NEMs demonstrated no cytotoxic effects on NIH3T3 cells even at higher concentration (1000 µg/mL), as confirmed by cell viability and fluorescent staining assays. In conclusion, this study suggests that the developed CS-UDA NEMs hold promise as potent antifungal agents with diverse potential applications.
Assuntos
Antifúngicos , Quitosana , Emulsões , Ácidos Graxos , Quitosana/química , Quitosana/farmacologia , Antifúngicos/farmacologia , Antifúngicos/química , Emulsões/química , Ácidos Graxos/química , Camundongos , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Testes de Sensibilidade Microbiana , Nanopartículas/química , Células NIH 3T3 , Antioxidantes/farmacologia , Antioxidantes/química , Candida albicans/efeitos dos fármacosRESUMO
The increasing prevalence of endocrine-disrupting chemicals (EDCs) in our environment is a growing concern, with numerous studies highlighting their adverse effects on the human endocrine system. Among the EDCs, estrogenic endocrine-disrupting chemicals (eEDCs) are exogenous compounds that perturb estrogenic hormone function by interfering with estrogen receptor (ER) homo (α/α, ß/ß) or hetero (α/ß) dimerization. To date, a comprehensive screening approach for eEDCs affecting all ER dimer forms in live cells is lacking. Here, we developed ER dimerization-detecting biosensors (ERDDBs), based on bioluminescence resonance energy transfer, for dimerization detection and rapid eEDC identification. To enhance the performance of these biosensors, we determined optimal donor and acceptor locations using computational analysis. Additionally, employing HaloTag as the acceptor and incorporating the P2A peptide as a linker yielded the highest sensitivity among the prototypes. We also established stable cell lines to screen potential ER dimerization inducers among estrogen analogs (EAs). The EAs were categorized through cross-comparison of ER dimer responses, utilizing EC values derived from a standard curve established with 17ß-estradiol. We successfully classified 26 of 72 EAs, identifying which ER dimerization types they induce. Overall, our study underscores the effectiveness of the optimized ERDDB for detecting ER dimerization and its applicability in screening and identifying eEDCs.
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Bacterial infections can lead to the formation of chronic wounds and delay the wound-healing process. Therefore, it is important to explore safe and efficient antimicrobial agents that have wound-healing and biocompatible properties. In this study, novel starch-fabricated silver nitroprusside nanoparticles (S-AgNP NPs) were prepared for biocompatible wound-healing applications. The study showed that S-AgNP NPs are spherical, with an average size of 356 ± 22.28 d. nm and zeta potential of -27.8 ± 2.80 mV, respectively. Furthermore, the FTIR and XRD results showed that S-AgNP NPs have functional groups and crystal structures from the silver nitroprusside nanoparticles (AgNP NPs) and starch. Additionally, S-AgNP NPs showed excellent bacterial and biofilm inhibition on B. cereus (15.6 µg/mL), L. monocytogenes (15.6 µg/mL), S. aureus (31.3 µg/mL), E. coli (31.3 µg/mL) and S. enterica (62.5 µg/mL). Moreover, S-AgNP NPs promoted cell migration and proliferation at a concentration of 62.5 µg/mL compared to AgNP NPs. Meanwhile, S-AgNP NPs had good biocompatibility and low cytotoxicity compared to AgNP NPs. Therefore, this study provided new ideas for the development of wound-healing agents with bacteriostatic properties in chronic wounds.
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Polysaccharides are abundantly present in fungi and are gaining recognition for their exceptional bioactivities. Hence, the present study aimed to extract intracellular polysaccharides (IPS-1 and IPS-2) from the endophytic Penicillium radiatolobatum and compare their physicochemical and bioactive attributes. The monosaccharide composition analysis revealed the existence of galactose, glucose, and mannose in both the IPS, while a trace amount of xylose was found in IPS-1. Further, FT-IR, 1H NMR, and 13C NMR analysis suggested that the IPS-2 was mainly composed of the ß-(1â4)-D-Galactose and ß-(1â4)-D-Glucose as the main chain, with the ß-(1â6)-D-mannose as branched chains. Compared to IPS-1, the IPS-2 showed higher antioxidant activities with an IC50 value of 108 ± 2.5 µg/mL, 272 ± 4.0 µg/mL, and 760 ± 5.0 µg/mL for ABTS+ scavenging, DPPH radical scavenging, and ferric reducing power, respectively. In addition, the IPS-2 inhibited the viability of prostate cancer (PC-3) cells (IC50; 435 ± 3.0 µg/mL) via apoptosis associated with mitochondrial membrane potential collapse and altered morphological features, which was revealed by cellular staining and flow cytometric analysis. Moreover, no apparent cytotoxic effects were seen in IPS-2-treated (1000 µg/mL) non-cancerous cells (HEK-293 and NIH3T3). Overall, the findings of this study suggest that P. radiatolobatum could be a potent source of polysaccharides with promising antioxidant and anticancer activity.
Assuntos
Antioxidantes , Penicillium , Animais , Camundongos , Humanos , Antioxidantes/farmacologia , Antioxidantes/química , Espectroscopia de Infravermelho com Transformada de Fourier , Células HEK293 , Células NIH 3T3 , Polissacarídeos/farmacologia , Polissacarídeos/química , Glucose/química , Manose/química , Galactose/químicaRESUMO
Tauopathies are neurodegenerative diseases characterized by abnormal conformational changes in tau protein. Early hyperphosphorylation-induced conformational changes are considered a hallmark of tauopathy, but real-time tracking methods are lacking. Here, we present two novel fluorescence resonance energy transfer (FRET)-based tau biosensors that detect such changes with high spatiotemporal resolution at the single-cell level. The TAUCON biosensor measures instantaneous conformational changes in hyperphosphorylated tau within 20 min, while the TAUCOM biosensor detects changes in the paper-clip structure of microtubule-associated tau. Our biosensors provide faster and more precise detection than conventional methods and can serve as valuable tools for investigating the initial causes, mechanisms, progression, and treatment of tauopathies.
Assuntos
Técnicas Biossensoriais , Doenças Neurodegenerativas , Tauopatias , Humanos , Proteínas tau/metabolismo , Transferência Ressonante de Energia de Fluorescência/métodos , Tauopatias/diagnóstico , Tauopatias/metabolismoRESUMO
In this study, two extracellular polysaccharides (TEPS1 and TEPS2) were isolated from the endophytic fungus (Talaromyces purpureogenus) and purified by DEAE-Sepharose Fast Flow column using NaCl as gradient eluent. The HPLC analysis displayed that TEPS1 was composed of mannose (38.70%), ribose (25.02%), glucose (19.34%), and galactose (16.94%) while the TEPS2 composed by mannose (100%). The NMR results indicated that TEPS1 exhibited α-glycosidic configurations. The both polysaccharides, TEPS1 and TEPS2 were exhibited a good antioxidant activity in terms of DPPH, ABTS, and â¢OH scavenging. However, TEPS1 showed a higher antioxidant activity than TEPS2. The IC50 of TEPS1 were 32.16, 192.57, and 54.67 µg·mL-1, for DPPH, ABTS, and â¢OH radical scavenging, respectively. Furthermore, TEPS1 showed the high cellular antioxidant and wound healing activity in the human embryonic kidney (HEK293) cell line. Overall, these two polysaccharides were promising in antioxidant activity.
Assuntos
Antioxidantes , Talaromyces , Humanos , Antioxidantes/química , Manose , Células HEK293 , Talaromyces/metabolismo , Polissacarídeos/química , CicatrizaçãoRESUMO
This work aimed to investigate the effect of EPS (extracellular polysaccharide) of Weissella cibaria as a prebiotic to promote the growth and antibacterial properties of Lactobacillus rhamnosus. The morphological, growth behavior, and antibacterial properties of L. rhamnosus were determined in MRSB (de Man Rogosa Sharpe broth) supplemented with different concentrations of EPS (0.1-2%). The results revealed that the incorporation of the EPS (2%) in MRSA improved the bacterial growth in terms of colony-forming unit (CFU, 0.7 × 105 CFU/mL) compared to L. rhamnosus grown in bare MRSA. The SEM observation revealed that EPS incorporation in the MRSB culture media does not affect the morphological properties of L. rhamnosus. Moreover, it was confirmed that the extract of probiotics cultured in MRSA supplemented with EPS (2%) was exhibited strong antibacterial and antibiofilm activity against targeted pathogens. This L. rhamnosus extract was found to be biocompatible evidanced by erythrocyte hemolysis assay. These results confirmed that EPS regulates the growth of probiotics, resists pathogen infection, and biocompatibility.
Assuntos
Lacticaseibacillus rhamnosus , Probióticos , Weissella , Humanos , Prebióticos , Lactobacillus/metabolismo , Weissella/metabolismo , Antibacterianos/farmacologiaRESUMO
A total of sixteen bacterial strains were isolated and identified from the fourteen types of Korean fermented foods that were evaluated for their in vitro probiotic potentials. The results showed the highest survivability for Bacillus sp. compared to Lactobacillus sp. in simulated gastric pH, and it was found to be maximum for B. inaquosorum KNUAS016 (8.25 ± 0.08 log10 CFU/mL) and minimum for L. sakei KNUAS019 (0.8 ± 0.02 log10 CFU/mL) at 3 h of incubation. Furthermore, B. inaquosorum KNUAS016 and L. brevis KNUAS017 also had the highest survival rates of 6.86 ± 0.02 and 5.37 ± 0.01 log10 CFU/mL, respectively, in a simulated intestinal fluid condition at 4 h of incubation. The percentage of autoaggregation at 6 h for L. sakei KNUAS019 (66.55 ± 0.33%), B. tequilensis KNUAS015 (64.56 ± 0.14%), and B. inaquosorum KNUAS016 (61.63 ± 0.19%) was >60%, whereas it was lower for L. brevis KNUAS017 (29.98 ± 0.09%). Additionally, B. subtilis KNUAS003 showed higher coaggregation at 63.84 ± 0.19% while B. proteolyticus KNUAS001 found at 30.02 ± 0.33%. Among them, Lactobacillus sp. showed the best non-hemolytic activity. The highest DPPH and ABTS radical scavenging activity was observed in L. sakei KNUAS019 (58.25% and 71.88%). The cell-free supernatant of Lactobacillus sp. considerably inhibited pathogenic growth, while the cell-free supernatant of Bacillus sp. was moderately inhibited when incubated for 24 h. However, the overall results found that B. subtilis KNUAS003, B. proteolyticus KNUAS012, L. brevis KNUAS017, L. graminis KNUAS018, and L. sakei KNUAS019 were recognized as potential probiotics through different functional and toxicity assessments.
Assuntos
Alimentos Fermentados , Probióticos , Antibacterianos/farmacologia , Antioxidantes/farmacologia , Lactobacillus , Probióticos/farmacologia , República da CoreiaRESUMO
Antimicrobial resistance (AMR) causes global consequences through increased mortality and economic loss. Antimicrobial drugs including nanomaterials are an emerging environmental impact. Hence, this work aimed to synthesize and characterize the titanium dioxide nanoparticles (TiO2 NPs) using the aqueous extract of endophytic fungus Paraconiothyrium brasiliense (Pb) for enhancing the antibacterial efficiency of existing standard antibiotics at minimum concentration. The FTIR and XRD results confirmed the capping of functional molecules and the crystalline nature of Pb-TiO2 NPs. The spherical-shaped TiO2 NPs with the size of 57.39 ± 13.65 nm were found in TEM analysis. The average hydrodynamic size (68.43 ± 1.49 d. nm) and the zeta potential (-19.6 ± 1.49 mV) was confirmed the stability of Pb-TiO2 NPs. Antibacterial studies revealed that bare Pb-TiO2 NPs (20 µg/mL) did not exhibit significant antibacterial activity while combination of TCH + Pb-TiO2 NPs considerably increased the inhibition of E. coli biofilm evidenced by CLSM and SEM analysis. Further, Pb-TiO2 NPs (100 µg/mL) were found to be moderately toxic to cell line (NIH3T3), red blood cells (RBC), and egg embryos. Hence, this study concluded that <50 µg/mL of TiO2 NPs can be mixed with antibiotics for enhanced antibacterial application thereby minimizing the AMR and the environmental toxicity.
Assuntos
Anti-Infecciosos , Nanopartículas Metálicas , Nanopartículas , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Ascomicetos , Biofilmes , Escherichia coli , Chumbo , Nanopartículas Metálicas/química , Nanopartículas Metálicas/toxicidade , Camundongos , Células NIH 3T3 , Nanopartículas/química , Nanopartículas/toxicidade , Titânio/química , Titânio/toxicidadeRESUMO
The present work reported synthesis, characterization, and biocompatibility of starch encapsulated silver nanoparticles (St-PF-AgNPs) and their antibacterial activity. The synthesis of St-PF-AgNPs involved in two steps: (i) synthesis of the biogenic silver nanoparticles using the fungal extracts (PF-AgNPs); and, (ii) encapsulation of starch in PF-AgNPs (St-PF-AgNPs). The surface plasmon resonance was found at 420 nm for the PF-AgNPs while it was at 260 and 420 nm for the St-PF-AgNPs. FTIR spectrum demonstrated the capping and encapsulation of the fungal extracts and starch in PF-AgNPs and St-PF-AgNPs. The XRD and TEM-EDS confirmed the crystalline nature, spherical-shaped , and polydispersed- PF-AgNPs and St-PF-AgNPs with strong signals of Ag. The St-PF-AgNPs showed a Z-average size of 115.2 d.nm and zeta potential of -17.8 (mV) as indicated by DLS and zeta potentials. The cytotoxicity results demonstrated higher toxicity of PF-AgNPs than St-PF-AgNPs in HEK293 cells. The antibacterial activity of St-PF-AgNPs were higher than PF-AgNPs in S. aureus. Overall, this work concluded that the starch encapsulation significantly increased the antibacterial activity of PF-AgNPs and this opens a new avenue for the treatment of bacterial infections through the sustained release of PF-AgNPs to target pathogenic bacterial cells.