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1.
Sci Rep ; 14(1): 12651, 2024 06 02.
Article in English | MEDLINE | ID: mdl-38825618

ABSTRACT

Effective disinfection methods are crucial in the cold chain transportation process of food due to the specificity of temperature and the diversity of contaminated flora. The objective of this study was to investigate the sanitizing effect of different disinfectants on various fungi at - 20 °C to achieve accurate disinfection of diverse bacterial populations. Peracetic acid, hydrogen peroxide, and potassium bisulfate were selected as low-temperature disinfectants and were combined with antifreeze. The sanitizing effect of these cryogenic disinfectants on pathogens such as Bacillus subtilis black variant spores (ATCC9372), Staphylococcus aureus (ATCC 6538), Candida albicans (ATCC 10231), Escherichia coli (8099), and poliovirus (PV-1) was sequentially verified by bactericidal and virus inactivation experiments. After a specified time of disinfection, a neutralizing agent was used to halt the sanitizing process. The study demonstrates that different disinfectants exhibit selective effects during the low-temperature disinfection process. Peracetic acid, hydrogen peroxide, and potassium monopersulfate are suitable for the low-temperature environmental disinfection of bacterial propagules, viruses, and fungal contaminants. However, for microorganisms with strong resistance to spores, a low-temperature disinfectant based on peracetic acid should be chosen for effective disinfection treatment. Our results provide a valuable reference for selecting appropriate disinfectants to sanitize various potential pathogens in the future.


Subject(s)
Cold Temperature , Disinfectants , Disinfection , Hydrogen Peroxide , Peracetic Acid , Disinfectants/pharmacology , Disinfection/methods , Hydrogen Peroxide/pharmacology , Peracetic Acid/pharmacology , Sulfates/pharmacology , Bacillus subtilis/drug effects , Potassium Compounds/pharmacology , Staphylococcus aureus/drug effects , Candida albicans/drug effects , Escherichia coli/drug effects , Poliovirus/drug effects
2.
Front Cell Infect Microbiol ; 14: 1375872, 2024.
Article in English | MEDLINE | ID: mdl-38846355

ABSTRACT

Introduction: Pseudomonas aeruginosa is notorious for its multidrug resistance and its involvement in hospital-acquired infections. In this study, 20 bacterial strains isolated from soil samples near the Hindan River in Ghaziabad, India, were investigated for their biochemical and morphological characteristics, with a focus on identifying strains with exceptional drug resistance and pyocyanin production. Methods: The isolated bacterial strains were subjected to biochemical and morphological analyses to characterize their properties, with a particular emphasis on exopolysaccharide production. Strain GZB16/CEES1, exhibiting remarkable drug resistance and pyocyanin production. Biochemical and molecular analyses, including sequencing of its 16S rRNA gene (accession number LN735036.1), plasmid-curing assays, and estimation of plasmid size, were conducted to elucidate its drug resistance mechanisms and further pyocynin based target the Candida albicans Strain GZB16/CEES1 demonstrated 100% resistance to various antibiotics used in the investigation, with plasmid-curing assays, suggesting plasmid-based resistance gene transmission. The plasmid in GZB16/CEES1 was estimated to be approximately 24 kb in size. The study focused on P. aeruginosa's pyocyanin production, revealing its association with anticandidal activity. The minimum inhibitory concentration (MIC) of the bacterial extract against Candida albicans was 50 µg/ml, with a slightly lower pyocyanin-based MIC of 38.5 µg/ml. Scanning electron microscopy illustrated direct interactions between P. aeruginosa strains and Candida albicans cells, leading to the destruction of the latter. Discussion: These findings underscore the potential of P. aeruginosa in understanding microbial interactions and developing strategies to combat fungal infections. The study highlights the importance of investigating bacterial-fungal interactions and the role of pyocyanin in antimicrobial activity. Further research in this area could lead to the development of novel therapeutic approaches for combating multidrug-resistant infections.


Subject(s)
Antifungal Agents , Candida albicans , Drug Resistance, Multiple, Bacterial , Microbial Sensitivity Tests , Plasmids , Pseudomonas aeruginosa , Pyocyanine , RNA, Ribosomal, 16S , Soil Microbiology , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/genetics , Pseudomonas aeruginosa/metabolism , Pyocyanine/metabolism , Drug Resistance, Multiple, Bacterial/genetics , Antifungal Agents/pharmacology , Candida albicans/drug effects , Candida albicans/genetics , Candida albicans/growth & development , RNA, Ribosomal, 16S/genetics , India , Plasmids/genetics , Anti-Bacterial Agents/pharmacology , Antibiosis
3.
Microb Cell Fact ; 23(1): 161, 2024 May 31.
Article in English | MEDLINE | ID: mdl-38822407

ABSTRACT

Multi resistant fungi are on the rise, and our arsenal compounds are limited to few choices in the market such as polyenes, pyrimidine analogs, azoles, allylamines, and echinocandins. Although each of these drugs featured a unique mechanism, antifungal resistant strains did emerge and continued to arise against them worldwide. Moreover, the genetic variation between fungi and their host humans is small, which leads to significant challenges in new antifungal drug discovery. Endophytes are still an underexplored source of bioactive secondary metabolites. Many studies were conducted to isolate and screen endophytic pure compounds with efficacy against resistant yeasts and fungi; especially, Candida albicans, C. auris, Cryptococcus neoformans and Aspergillus fumigatus, which encouraged writing this review to critically analyze the chemical nature, potency, and fungal source of the isolated endophytic compounds as well as their novelty features and SAR when possible. Herein, we report a comprehensive list of around 320 assayed antifungal compounds against Candida albicans, C. auris, Cryptococcus neoformans and Aspergillus fumigatus in the period 1980-2024, the majority of which were isolated from fungi of orders Eurotiales and Hypocreales associated with terrestrial plants, probably due to the ease of laboratory cultivation of these strains. 46% of the reviewed compounds were active against C. albicans, 23% against C. neoformans, 29% against A. fumigatus and only 2% against C. auris. Coculturing was proved to be an effective technique to induce cryptic metabolites absent in other axenic cultures or host extract cultures, with Irperide as the most promising compounds MIC value 1 µg/mL. C. auris was susceptible to only persephacin and rubiginosin C. The latter showed potent inhibition against this recalcitrant strain in a non-fungicide way, which unveils the potential of fungal biofilm inhibition. Further development of culturing techniques and activation of silent metabolic pathways would be favorable to inspire the search for novel bioactive antifungals.


Subject(s)
Antifungal Agents , Endophytes , Antifungal Agents/pharmacology , Endophytes/metabolism , Humans , Microbial Sensitivity Tests , Cryptococcus neoformans/drug effects , Cryptococcus neoformans/metabolism , Fungi/drug effects , Fungi/metabolism , Aspergillus fumigatus/drug effects , Aspergillus fumigatus/metabolism , Candida albicans/drug effects
4.
AAPS PharmSciTech ; 25(5): 130, 2024 Jun 06.
Article in English | MEDLINE | ID: mdl-38844611

ABSTRACT

Naringenin (NRG) inhibits the fungal 17ß-hydroxysteroid dehydrogenase accountable for ergosterol synthesis in Candida albicans (C. albicans), a causative agent for cutaneous candidiasis. In present research, NRG was complexed with ZnO nanomaterial (NRG-Zn2+) to synthesize NRG-Zn2+ nanocomposites. The particle size and ζ-potential of NRG-Zn2+ nanocomposites were respectively estimated to be 180.33 ± 1.22-nm and - 3.92 ± 0.35-mV. In silico data predicted the greater affinity of NRG-Zn2+ nanocomposite for 14α-demethylase and ceramide in comparison to NRG alone. Later, NRG-Zn2+ nanocomposites solution was transformed in to naringenin-zinc oxide nanocomposites loaded chitosan gel (NRG-Zn-CS-Gel) with viscosity and firmness of 854806.7 ± 52386.43 cP and 698.27 ± 10.35 g, respectively. The ex-vivo skin permeation demonstrated 70.49 ± 5.22% skin retention, significantly greater (P < 0.05) than 44.48 ± 3.06% of naringenin loaded chitosan gel (NRG-CS-Gel) and 31.24 ± 3.28% of naringenin solution (NRG Solution). NRG-Zn-CS-Gel demonstrated 6.71 ± 0.84% permeation of NRG with a flux value of 0.046 ± 0.01-µg/cm2/h. The MIC50 of NRG-Zn-CS-Gel against C. albicans was estimated to be 0.156-µg/mL with FICI (fractional inhibitory concentration index) of 0.018 that consequently exhibited synergistic efficacy. Further, NRG-Zn-CS-Gel demonstrated superior antifungal efficacy in C. albicans induced cutaneous candidiasis infection in Balb/c mice. The fungal burden in NRG-Zn-CS-Gel treated group was 109 ± 25 CFU/mL, significantly lower (P < 0.05) than positive control (2260 ± 446 CFU/mL), naringenin loaded chitosan gel (NRG-CS-Gel; 928 ± 127 CFU/mL) and chitosan gel (CS-Gel; 2116 ± 186 CFU/mL) treated mice. Further, histopathology examination and cytokine profiling of TNF-α, IL-1ß and IL-10 revealed the healing of skin and inflammation associated with cutaneous candidiasis infection. In conclusion, NRG-Zn-CS-Gel may be a potential candidate for translating in to a clinical viable topical nanotherapeutic.


Subject(s)
Antifungal Agents , Candida albicans , Chitosan , Flavanones , Gels , Mice, Inbred BALB C , Nanocomposites , Zinc Oxide , Animals , Flavanones/administration & dosage , Flavanones/pharmacology , Mice , Candida albicans/drug effects , Chitosan/chemistry , Chitosan/administration & dosage , Nanocomposites/chemistry , Nanocomposites/administration & dosage , Antifungal Agents/administration & dosage , Antifungal Agents/pharmacology , Antifungal Agents/pharmacokinetics , Zinc Oxide/administration & dosage , Zinc Oxide/pharmacology , Zinc Oxide/chemistry , Drug Delivery Systems/methods , Skin/metabolism , Skin/drug effects , Skin/microbiology , Candidiasis/drug therapy , Polymers/chemistry , Skin Absorption/drug effects , Particle Size , Administration, Cutaneous
5.
Sci Rep ; 14(1): 12997, 2024 06 06.
Article in English | MEDLINE | ID: mdl-38844768

ABSTRACT

Herbal medicine combined with nanoparticles has caught much interest in clinical dental practice, yet the incorporation of chitosan with Salvadora persica (S. persica) extract as an oral care product has not been explored. The aim of this study was to evaluate the combined effectiveness of Salvadora persica(S. persica) and Chitosan nanoparticles (ChNPs) against oropharyngeal microorganisms. Agar well diffusion, minimum inhibitory concentration, and minimal lethal concentration assays were used to assess the antimicrobial activity of different concentrations of ethanolic extracts of S. persica and ChNPs against selected fungal strains, Gram-positive, and Gram-negative bacteria. A mixture of 10% S. persica and 0.5% ChNPs was prepared (SChNPs) and its synergistic effect against the tested microbes was evaluated. Furthermore, the strain that was considered most sensitive was subjected to a 24-h treatment with SChNPs mixture; and examined using SEM, FT-IR and GC-MS analysis. S. persica extract and ChNPs exhibited concentration-dependent antimicrobial activities against all tested strains. S. persica extract and ChNPs at 10% were most effective against S. pneumoni, K. pneumoni, and C. albicans. SEM images confirmed the synergistic effect of the SChNPs mixture, revealing S. pneumonia cells with increased irregularity and higher cell lysis compared to the individual solutions. GC-MS and FT-IR analysis of SChNPs showed many active antimicrobial phytocompounds and some additional peaks, respectively. The synergy of the mixture of SChNPs in the form of mouth-rinsing solutions can be a promising approach for the control of oropharyngeal microbes that are implicated in viral secondary bacterial infections.


Subject(s)
Chitosan , Drug Synergism , Microbial Sensitivity Tests , Nanoparticles , Plant Extracts , Salvadoraceae , Chitosan/pharmacology , Chitosan/chemistry , Nanoparticles/chemistry , Plant Extracts/pharmacology , Plant Extracts/chemistry , Salvadoraceae/chemistry , Oropharynx/microbiology , Oropharynx/drug effects , Anti-Infective Agents/pharmacology , Anti-Infective Agents/chemistry , Candida albicans/drug effects , Humans , Spectroscopy, Fourier Transform Infrared
6.
Int J Nanomedicine ; 19: 4941-4956, 2024.
Article in English | MEDLINE | ID: mdl-38828194

ABSTRACT

Background: Due to its prevalence, recurrence, and the emergence of drug-resistance, Candida vaginitis significantly impacts the well-being of women. Although cinnamon essential oil (CEO) possesses antifungal activity, its hydrophobic properties limit its clinical application. Purpose: To overcome this challenge, a nanoemulsification technology was employed to prepare cinnamon essential oil-nanoemulsion (CEO@NE), and its therapeutic efficacy and action mechanism for Candida vaginitis was investigated in vivo and in vitro. Materials and Methods: CEO@NE, composed of 4% CEO, 78% distilled water, and 18% Tween 80, was prepared by ultrasonic nanoemulsification. The physical properties, anti-Candida activity, cytotoxicity, immunomodulatory potential and storage stability of CEO@NE were explored. Subsequently, the effect of intravaginal CEO@NE treatment on Candida vaginitis was investigated in mice. To comprehend the possible mechanism of CEO@NE, an analysis was conducted to ascertain the production of intracellular reactive oxygen species (ROS) in C. albicans. Results: CEO@NE, with the droplet size less than 100 nm and robust storage stability for up to 8 weeks, exhibited comparable anti-Candida activity with CEO. CEO@NE at the concentration lower than 400 µg/mL had no cytotoxic and immunomodulatory effects on murine splenocytes. Intravaginal treatment of CEO@NE (400 µg/mL, 20 µL/day/mouse for 5 consecutive days) curbed Candida colonization, ameliorated histopathological changes, and suppressed inflammatory cytokine production in mice intravaginally challenged with C. albicans. Notably, this treatment preserved the density of vaginal lactic acid bacteria (LAB) crucial for vaginal health. Co-culturing C. albicans with CEO@NE revealed concentration-dependent augmentation of intracellular ROS generation and ensuing cell death. In addition, co-culturing LPS-stimulated murine splenocytes with CEO@NE yielded a decrease in the generation of cytokines. Conclusion: This discovery provides insight into the conceivable antifungal and anti-inflammatory mechanisms of CEO@NE to tackle Candida vaginitis. CEO@NE offers a promising avenue to address the limitations of current treatments, providing novel strategy for treating Candida vaginitis.


Subject(s)
Antifungal Agents , Candida albicans , Candidiasis, Vulvovaginal , Cinnamomum zeylanicum , Emulsions , Oils, Volatile , Female , Animals , Oils, Volatile/chemistry , Oils, Volatile/pharmacology , Oils, Volatile/administration & dosage , Candidiasis, Vulvovaginal/drug therapy , Candida albicans/drug effects , Antifungal Agents/pharmacology , Antifungal Agents/chemistry , Antifungal Agents/administration & dosage , Mice , Administration, Intravaginal , Cinnamomum zeylanicum/chemistry , Emulsions/chemistry , Reactive Oxygen Species/metabolism , Humans , Nanoparticles/chemistry , Mice, Inbred BALB C
7.
Saudi Med J ; 45(6): 606-616, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38830660

ABSTRACT

OBJECTIVES: To assess the risk variables related to the types of candidemia for each patient, who was admitted into the intensive care unit regardless of the patient with or without complete diagnosis of COVID-19, during the period of March 2019 to December 2022. METHODS: The evaluation comparison of demographic and clinical data of COVID-19 positive and negative patients with candidemia confirmed in blood, 113 cases were assessed. Variables such as gender, age, age of hospitalization, history of hospitalization, concurrently infection, The acute physiology and chronic health evaluation-II scores, comorbidity checking, intubation, central venous catheter use, parenteral nutrition use, steroid use, antibiotic use, lymphopenia, and laboratory variables were evaluated. Candida species distribution, antifungal susceptibility in blood culture were determined. RESULTS: Coronavirus disease-19 was present in 62.8% of cases confirmed candidemia, and these cases were significantly different from COVID-19 negative cases. Significance was found in more intubation, central venous catheter use, parenteral nutrition, and steroid therapy in Group 2. There was no significance with species distribution and associated infection. In total, COVID-19 positive had higher hemoglobin, aspartate aminotransferase, alanine transaminase, and white blood cell levels, which may be associated with the possibility of revealing and controlling candidemia. CONCLUSION: Candida albicans and Candida Parapsilosis (C. parapsilosis) are the species seen in infected COVID-19 patients, while C. parapsilosis and Candida tropicalis are found in non-COVID-19 ones. Risk factors were intubation, parenteral nutrition, central venous catheter, and steroid in the COVID-19 group.


Subject(s)
COVID-19 , Candida , Candidemia , Intensive Care Units , Humans , Candidemia/epidemiology , Risk Factors , Male , Female , Intensive Care Units/statistics & numerical data , COVID-19/complications , COVID-19/epidemiology , Middle Aged , Candida/isolation & purification , Aged , Adult , Parenteral Nutrition , Candida albicans/isolation & purification , Antifungal Agents/therapeutic use , SARS-CoV-2 , Candida tropicalis/isolation & purification
8.
Curr Microbiol ; 81(7): 213, 2024 Jun 07.
Article in English | MEDLINE | ID: mdl-38847863

ABSTRACT

The antimalarial drug Mefloquine has demonstrated antifungal activity against growth and virulence factors of Candida albicans. The current study focused on the identification of Mefloquine's mode of action in C. albicans by performing cell susceptibility assay, biofilm assay, live and dead assay, propidium iodide uptake assay, ergosterol quantification assay, cell cycle study, and gene expression studies by RT-PCR. Mefloquine inhibited the virulence factors in C. albicans, such as germ tube formation and biofilm formation at 0.125 and 1 mg/ml, respectively. Mefloquine-treated cells showed a decrease in the quantity of ergosterol content of cell membrane in a concentration-dependent manner. Mefloquine (0.25 mg/ml) arrested C. albicans cells at the G2/M phase and S phase of the cell cycle thereby preventing the progression of the normal yeast cell cycle. ROS level was measured to find out oxidative stress in C. albicans in the presence of mefloquine. The study revealed that, mefloquine was found to enhance the ROS level and subsequently oxidative stress. Gene expression studies revealed that mefloquine treatment upregulates the expressions of SOD1, SOD2, and CAT1 genes in C. albicans. In vivo, the antifungal efficacy of mefloquine was confirmed in mice for systemic candidiasis and it was found that there was a decrease in the pathogenesis of C. albicans after the treatment of mefloquine in mice. In conclusion, mefloquine can be used as a repurposed drug as an alternative drug against Candidiasis.


Subject(s)
Antifungal Agents , Candida albicans , Candidiasis , Mefloquine , Virulence Factors , Antifungal Agents/pharmacology , Candida albicans/drug effects , Candida albicans/genetics , Candida albicans/pathogenicity , Candida albicans/growth & development , Animals , Mefloquine/pharmacology , Mice , Virulence Factors/genetics , Virulence Factors/metabolism , Candidiasis/microbiology , Candidiasis/drug therapy , Biofilms/drug effects , Biofilms/growth & development , Reactive Oxygen Species/metabolism , Microbial Sensitivity Tests , Oxidative Stress/drug effects , Cell Cycle/drug effects , Superoxide Dismutase-1/genetics , Superoxide Dismutase-1/metabolism , Ergosterol/metabolism , Fungal Proteins/genetics , Fungal Proteins/metabolism
9.
Invest Ophthalmol Vis Sci ; 65(6): 13, 2024 Jun 03.
Article in English | MEDLINE | ID: mdl-38848078

ABSTRACT

Purpose: Fungal keratitis (FK) is an invasive corneal infection associated with significant risk to vision. Although the cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) signaling pathway has been recognized for its role in defending against viral infections, its involvement in FK still remains largely unclear. This study sought to elucidate the contribution of the cGAS/STING signaling pathway to the pathogenesis of FK. Methods: The expression of cGAS/STING signaling components was assessed in a murine model of Candida albicans keratitis through RNA sequencing, western blot analysis, immunofluorescence staining, and real-time PCR. Both genetic (utilizing Sting1gt/gt mice) and pharmacological (using C176) interventions were employed to inhibit STING activity, allowing for the evaluation of resultant pathogenic alterations in FK using slit-lamp examination, clinical scoring, hematoxylin and eosin (H&E) staining, fungal culture, and RNA sequencing. Subconjunctival administration of the NOD-like receptor protein 3 (NLRP3) inflammasome inhibitor MCC950 was performed to evaluate FK manifestations following STING activity blockade. Furthermore, the impact of the STING agonist diABZI on FK progression was investigated. Results: Compared to uninfected corneas, those infected with C. albicans exhibited increased expression of cGAS/STING signaling components, as well as its elevated activity. Inhibiting cGAS/STING signaling exacerbated the advancement of FK, as evidenced by elevated clinical scores, augmented fungal load, and heightened inflammatory response, including NLRP3 inflammasome activation and pyroptosis. Pharmacological inhibition of the NLRP3 inflammasome effectively mitigated the exacerbated FK by suppressing STING activity. Conversely, pre-activation of STING exacerbated FK progression compared to the PBS control, characterized by increased fungal burden and reinforced inflammatory infiltration. Conclusions: This study demonstrates the essential role of the cGAS/STING signaling pathway in FK pathogenesis and highlights the necessity of its proper activation for the host against FK.


Subject(s)
Candida albicans , Candidiasis , Disease Models, Animal , Eye Infections, Fungal , Membrane Proteins , Nucleotidyltransferases , Signal Transduction , Animals , Membrane Proteins/metabolism , Membrane Proteins/genetics , Nucleotidyltransferases/metabolism , Nucleotidyltransferases/genetics , Eye Infections, Fungal/microbiology , Eye Infections, Fungal/metabolism , Mice , Candida albicans/physiology , Candidiasis/microbiology , Candidiasis/metabolism , Mice, Inbred C57BL , Real-Time Polymerase Chain Reaction , Keratitis/microbiology , Keratitis/metabolism , Blotting, Western , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/genetics , Female , Corneal Ulcer/microbiology , Corneal Ulcer/metabolism , Inflammasomes/metabolism
10.
Life Sci Alliance ; 7(7)2024 Jul.
Article in English | MEDLINE | ID: mdl-38719750

ABSTRACT

Celiac disease (CD) is an autoimmune enteropathy resulting from an interaction between diet, genome, and immunity. Although many patients respond to a gluten-free diet, in a substantive number of individuals, the intestinal injury persists. Thus, other factors might amplify the ongoing inflammation. Candida albicans is a commensal fungus that is well adapted to the intestinal life. However, specific conditions increase Candida pathogenicity. The hypothesis that Candida may be a trigger in CD has been proposed after the observation of similarity between a fungal wall component and two CD-related gliadin T-cell epitopes. However, despite being implicated in intestinal disorders, Candida may also protect against immune pathologies highlighting a more intriguing role in the gut. Herein, we postulated that a state of chronic inflammation associated with microbial dysbiosis and leaky gut are favorable conditions that promote C. albicans pathogenicity eventually contributing to CD pathology via a mast cells (MC)-IL-9 axis. However, the restoration of immune and microbial homeostasis promotes a beneficial C. albicans-MC cross-talk favoring the attenuation of CD pathology to alleviate CD pathology and symptoms.


Subject(s)
Candida albicans , Celiac Disease , Homeostasis , Mast Cells , Celiac Disease/immunology , Celiac Disease/microbiology , Celiac Disease/metabolism , Humans , Candida albicans/pathogenicity , Candida albicans/immunology , Mast Cells/immunology , Mast Cells/metabolism , Gastrointestinal Microbiome/immunology , Dysbiosis/immunology , Candidiasis/immunology , Candidiasis/microbiology , Animals , Candida/pathogenicity , Candida/immunology , Intestinal Mucosa/microbiology , Intestinal Mucosa/immunology , Intestinal Mucosa/metabolism
11.
AAPS PharmSciTech ; 25(5): 106, 2024 May 09.
Article in English | MEDLINE | ID: mdl-38724834

ABSTRACT

The primary factor underlying the virulence of Candida albicans is its capacity to form biofilms, which in turn leads to recurrent complications. Over-the-counter antifungal treatments have proven ineffective in eliminating fungal biofilms and the inflammatory cytokines produced during fungal infections. Chitosan nanoparticles offer broad and versatile therapeutic potential as both antifungal agents and carriers for antifungal drugs to combat biofilm-associated Candida infections. In our study, we endeavoured to develop chitosan nanoparticles utilising chitosan and the antifungal crosslinker phytic acid targeting C. albicans. Phytic acid, known for its potent antifungal and anti-inflammatory properties, efficiently crosslinks with chitosan. The nanoparticles were synthesised using the ionic gelation technique and subjected to analyses including Fourier transform infrared spectroscopy, dynamic light scattering, and zeta potential analysis. The synthesised nanoparticles exhibited dimensions with a diameter (Dh) of 103 ± 3.9 nm, polydispersity index (PDI) of 0.33, and zeta potential (ZP) of 37 ± 2.5 mV. These nanoparticles demonstrated an antifungal effect with a minimum inhibitory concentration (MIC) of 140 ± 2.2 µg/mL, maintaining cell viability at approximately 90% of the MIC value and reducing cytokine levels. Additionally, the nanoparticles reduced ergosterol content and exhibited a 62% ± 1.2 reduction in biofilm susceptibility, as supported by colony-forming unit (CFU) and XTT assays-furthermore, treatment with nanoparticles reduced exopolysaccharide production and decreased secretion of aspartyl protease by C. albicans. Our findings suggest that the synthesised nanoparticles effectively combat Candida albicans infections. In vivo studies conducted on a mouse model of vaginal candidiasis confirmed the efficacy of the nanoparticles in combating fungal infections in vivo.


Subject(s)
Antifungal Agents , Biofilms , Candida albicans , Chitosan , Microbial Sensitivity Tests , Nanoparticles , Phytic Acid , Chitosan/chemistry , Biofilms/drug effects , Nanoparticles/chemistry , Antifungal Agents/pharmacology , Antifungal Agents/administration & dosage , Animals , Candida albicans/drug effects , Mice , Microbial Sensitivity Tests/methods , Phytic Acid/pharmacology , Phytic Acid/administration & dosage , Phytic Acid/chemistry , Female , Candidiasis/drug therapy , Particle Size , Drug Carriers/chemistry , Cross-Linking Reagents/chemistry , Cytokines/metabolism
12.
Arch Microbiol ; 206(6): 251, 2024 May 10.
Article in English | MEDLINE | ID: mdl-38727840

ABSTRACT

The prevalence of Candida albicans infection has increased during the past few years, which contributes to the need for new, effective treatments due to the increasing concerns regarding antifungal drug toxicity and multidrug resistance. Butyl isothiocyanate (butylITC) is a glucosinolate derivative, and has shown a significant antifungal effect contrary to Candida albicans. Additionally, how butylITC affects the virulence traits of C. albicans and molecular mode of actions are not well known. Present study shows that at 17.36 mM concentration butylITC inhibit planktonic growth. butylITC initially slowed the hyphal transition at 0.542 mM concentration. butylITC hampered biofilm development, and inhibits biofilm formation at 17.36 mM concentration which was analysed using metabolic assay (XTT assay) and Scanning Electron Microscopy (SEM). In addition, it was noted that butylITC inhibits ergosterol biosynthesis. The permeability of cell membranes was enhanced by butylITC treatment. Moreover, butylITC arrests cells at S-phase and induces intracellular Reactive Oxygen Species (ROS) accumulation in C. albicans. The results suggest that butylITC may have a dual mode of action, inhibit virulence factors and modulate cellular processes like inhibit ergosterol biosynthesis, cell cycle arrest, induces ROS production which leads to cell death in C. albicans.


Subject(s)
Antifungal Agents , Biofilms , Candida albicans , Cell Membrane , Isothiocyanates , Oxidative Stress , Reactive Oxygen Species , Candida albicans/drug effects , Candida albicans/physiology , Biofilms/drug effects , Antifungal Agents/pharmacology , Isothiocyanates/pharmacology , Oxidative Stress/drug effects , Cell Membrane/drug effects , Cell Membrane/metabolism , Reactive Oxygen Species/metabolism , Microbial Sensitivity Tests , Cell Cycle/drug effects , Hyphae/drug effects , Hyphae/growth & development , Ergosterol/metabolism
13.
J Inorg Biochem ; 256: 112572, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38691971

ABSTRACT

Recognizing that metal ions play an important role in modifying the pharmacological properties of known organic-based drugs, the present manuscript addresses the complexation of the antifungal agent voriconazole (vcz) with the biologically relevant silver(I) ion as a strategy for the development of new antimycotics. The synthesized silver(I) complexes with vcz were characterized by mass spectrometry, IR, UV-Vis and NMR spectroscopy and single-crystal X-ray diffraction analysis. The crystallographic results showed that complexes {[Ag(vcz)(H2O)]CH3SO3}n (1), {[Ag(vcz)2]BF4}n (2) and {[Ag(vcz)2]PF6}n (3) have polymeric structures in the solid state, in which silver(I) ions have a distorted tetrahedral geometry. On the other hand, DFT calculations revealed that the investigated silver(I) complexes 1-3 in DMSO exist as linear [Ag(vcz-N2)(vcz-N19)]+ (1a), [Ag(vcz-N2)(vcz-N4)]+ (2a) and [Ag(vcz-N4)2]+ (3a) species, respectively. The evaluated complexes showed an enhanced anti-Candida activity compared to the parent drug with minimal inhibitory concentration (MIC) values in the range of 0.02-1.05 µM. In comparison with vcz, the corresponding silver(I) complexes showed better activity in prevention hyphae and biofilm formation of C. albicans, indicating that they could be considered as promising agents against Candida that significantly inhibit its virulence. Also, these complexes are much better inhibitors of ergosterol synthesis in the cell membrane of C. albicans at the concentration of 0.5 × MIC. This is also confirmed by a molecular docking, which revealed that complexes 1a - 3a showed better inhibitory activity than vcz against the sterol 14α-demethylase enzyme cytochrome P450 (CYP51B), which plays a crucial role in the formation of ergosterol.


Subject(s)
Antifungal Agents , Coordination Complexes , Microbial Sensitivity Tests , Silver , Voriconazole , Antifungal Agents/pharmacology , Antifungal Agents/chemistry , Antifungal Agents/chemical synthesis , Voriconazole/pharmacology , Voriconazole/chemistry , Silver/chemistry , Silver/pharmacology , Coordination Complexes/pharmacology , Coordination Complexes/chemistry , Coordination Complexes/chemical synthesis , Candida albicans/drug effects , Candida/drug effects , Crystallography, X-Ray
14.
Wiad Lek ; 77(3): 429-436, 2024.
Article in English | MEDLINE | ID: mdl-38691783

ABSTRACT

OBJECTIVE: Aim: The purpose of this study is to assess the impact of occupational hygiene procedures for microbiological and cytological contents of periodontal pockets. PATIENTS AND METHODS: Material and Methods: Cytological and microbiological content of the periodontal pockets before treatment and after professional hygiene procedures including scaling with hand instruments and root cementum polishing have been investigated in patients with periodontitis. RESULTS: Results: According to obtained data it can be resumed that in periodontitis patients with the depth of pockets 3-5,5 mm before professional hygiene all the pockets contain great number of Cocci, Spirochetes, Candida Albicans, Flagellated rods and Protozoa species. It was proved by revealing of small amount of Polymorphonuclear leukocytes with active phagocytosis. After scaling and planing of the roots, a decrease in the number of Protozoa and Candida Albicans was observed in 97% and 72% of the investigated cells, respectively. CONCLUSION: Conclusions: Cytological and microbiological content of periodontal pockets before treatment and after professional hygiene procedures including scaling and root planning testify to the level of local protective mechanisms, especially process of phagocytosis and virulence of microbial species in periodontal pockets.


Subject(s)
Periodontitis , Humans , Periodontitis/microbiology , Male , Female , Periodontal Pocket/microbiology , Middle Aged , Adult , Candida albicans/isolation & purification , Dental Scaling
15.
J Appl Oral Sci ; 32: e20230397, 2024.
Article in English | MEDLINE | ID: mdl-38695444

ABSTRACT

Specific products containing natural resources can contribute to the innovation of complete denture hygiene. OBJECTIVE: To conduct an in vitro evaluation of experimental dentifrices containing essential oils of Bowdichia virgilioides Kunth (BvK), Copaifera officinalis (Co), Eucalyptus citriodora (Ec), Melaleuca alternifolia (Ma) and Pinus strobus (Ps) at 1%. METHODOLOGY: The variables evaluated were organoleptic and physicochemical characteristics, abrasiveness (mechanical brushing machine) simulating 2.5 years, and microbial load (Colony Forming Units - CFU/mL), metabolic activity (XTT assay) and cell viability (Live/Dead® BacLight™ kit) of the multispecies biofilm (Streptococcus mutans: Sm, Staphylococcus aureus: Sa, Candida albicans: Ca and Candida glabrata: Cg). Specimens of heat-polymerized acrylic resins (n=256) (n=96 specimens for abrasiveness, n=72 for microbial load count, n=72 for biofilm metabolic activity, n=16 for cell viability and total biofilm quantification) with formed biofilm were divided into eight groups for manual brushing (20 seconds) with a dental brush and distilled water (NC: negative control), Trihydral (PC: positive control), placebo (Pl), BvK, Co, Ec, Ma or Ps. After brushing, the specimens were washed with PBS and immersed in Letheen Broth medium, and the suspension was sown in solid specific medium. The organoleptic characteristics were presented by descriptive analysis. The values of density, pH, consistency and viscosity were presented in a table. The data were analyzed with the Wald test in a generalized linear model, followed by the Kruskal-Wallis test, Dunn's test (mass change) and the Bonferroni test (UFC and XTT). The Wald test in Generalized Estimating Equations and the Bonferroni test were used to analyze cell viability. RESULTS: All dentifrices showed stable organoleptic characteristics and adequate physicochemical properties. CN, Ec, Ps, Pl and PC showed low abrasiveness. There was a significant difference between the groups (p<0.001) for microbial load, metabolic activity and biofilm viability. CONCLUSIONS: It was concluded that the BvK, Ec and Ps dentifrices are useful for cleaning complete dentures, as they have antimicrobial activity against biofilm. The dentifrices containing Bowdichia virgilioides Kunth showed medium abrasiveness and should be used with caution.


Subject(s)
Biofilms , Dentifrices , Denture, Complete , Materials Testing , Oils, Volatile , Biofilms/drug effects , Dentifrices/pharmacology , Dentifrices/chemistry , Oils, Volatile/pharmacology , Oils, Volatile/chemistry , Denture, Complete/microbiology , Time Factors , Reproducibility of Results , Toothbrushing , Colony Count, Microbial , Staphylococcus aureus/drug effects , Statistics, Nonparametric , Streptococcus mutans/drug effects , Analysis of Variance , Microbial Viability/drug effects , Candida albicans/drug effects , Reference Values , Acrylic Resins/chemistry , Acrylic Resins/pharmacology
16.
BMC Biotechnol ; 24(1): 28, 2024 May 04.
Article in English | MEDLINE | ID: mdl-38702622

ABSTRACT

Scientists know very little about the mechanisms underlying fish skin mucus, despite the fact that it is a component of the immune system. Fish skin mucus is an important component of defence against invasive infections. Recently, Fish skin and its mucus are gaining interest among immunologists. Characterization was done on the obtained silver nanoparticles Ag combined with Clarias gariepinus catfish epidermal mucus proteins (EMP-Ag-NPs) through UV-vis, FTIR, XRD, TEM, and SEM. Ag-NPs ranged in size from 4 to 20 nm, spherical in form and the angles were 38.10°, 44.20°, 64.40°, and 77.20°, Where wavelength change after formation of EMP-Ag-NPs as indicate of dark brown, the broad band recorded at wavelength at 391 nm. Additionally, the antimicrobial, antibiofilm and anticancer activities of EMP-Ag-NPs was assessed. The present results demonstrate high activity against unicellular fungi C. albicans, followed by E. faecalis. Antibiofilm results showed strong activity against both S. aureus and P. aeruginosa pathogens in a dose-dependent manner, without affecting planktonic cell growth. Also, cytotoxicity effect was investigated against normal cells (Vero), breast cancer cells (Mcf7) and hepatic carcinoma (HepG2) cell lines at concentrations (200-6.25 µg/mL) and current results showed highly anticancer effect of Ag-NPs at concentrations 100, 5 and 25 µg/mL exhibited rounding, shrinkage, deformation and granulation of Mcf7 and HepG2 with IC50 19.34 and 31.16 µg/mL respectively while Vero cells appeared rounded at concentration 50 µg/mL and normal shape at concentration 25, 12.5 and 6.25 µg/ml with IC50 35.85 µg/mL. This study evidence the potential efficacy of biologically generated Ag-NPs as a substitute medicinal agent against harmful microorganisms. Furthermore, it highlights their inhibitory effect on cancer cell lines.


Subject(s)
Biofilms , Catfishes , Metal Nanoparticles , Silver , Metal Nanoparticles/chemistry , Biofilms/drug effects , Biofilms/growth & development , Silver/chemistry , Silver/pharmacology , Animals , Humans , Mucus/metabolism , Antineoplastic Agents/pharmacology , Antineoplastic Agents/chemistry , Vero Cells , Fish Proteins/pharmacology , Fish Proteins/chemistry , Fish Proteins/metabolism , Chlorocebus aethiops , Cell Line, Tumor , Microbial Sensitivity Tests , Pseudomonas aeruginosa/drug effects , Anti-Infective Agents/pharmacology , Anti-Infective Agents/chemistry , Staphylococcus aureus/drug effects , Staphylococcus aureus/physiology , Candida albicans/drug effects , Epidermis/metabolism
17.
BMC Microbiol ; 24(1): 154, 2024 May 04.
Article in English | MEDLINE | ID: mdl-38704559

ABSTRACT

BACKGROUND: Side effects associated with antimicrobial drugs, as well as their high cost, have prompted a search for low-cost herbal medicinal substances with fewer side effects. These substances can be used as supplements to medicine or to strengthen their effects. The current study investigated the effect of oleuropein on the inhibition of fungal and bacterial biofilm in-vitro and at the molecular level. MATERIALS AND METHODS: In this experimental study, antimicrobial properties were evaluated using microbroth dilution method. The effect of oleuropein on the formation and eradication of biofilm was assessed on 96-well flat bottom microtiter plates and their effects were observed through scanning electron microscopy (SEM). Its effect on key genes (Hwp1, Als3, Epa1, Epa6, LuxS, Pfs) involved in biofilm formation was investigated using the quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) method. RESULTS: The minimum inhibitory concentration (MIC) and minimum fungicidal/bactericidal concentration (MFC/MBC) for oleuropein were found to be 65 mg/ml and 130 mg/ml, respectively. Oleuropein significantly inhibited biofilm formation at MIC/2 (32.5 mg/ml), MIC/4 (16.25 mg/ml), MIC/8 (8.125 mg/ml) and MIC/16 (4.062 mg/ml) (p < 0.0001). The anti-biofilm effect of oleuropein was confirmed by SEM. RT-qPCR indicated significant down regulation of expression genes involved in biofilm formation in Candida albicans (Hwp1, Als3) and Candida glabrata (Epa1, Epa6) as well as Escherichia coli (LuxS, Pfs) genes after culture with a MIC/2 of oleuropein (p < 0.0001). CONCLUSIONS: The results indicate that oleuropein has antifungal and antibacterial properties that enable it to inhibit or destroy the formation of fungal and bacterial biofilm.


Subject(s)
Antifungal Agents , Biofilms , Candida albicans , Candida glabrata , Escherichia coli , Fluconazole , Iridoid Glucosides , Iridoids , Microbial Sensitivity Tests , Biofilms/drug effects , Biofilms/growth & development , Iridoid Glucosides/pharmacology , Candida glabrata/drug effects , Candida glabrata/physiology , Candida glabrata/genetics , Candida albicans/drug effects , Candida albicans/genetics , Candida albicans/physiology , Escherichia coli/drug effects , Escherichia coli/genetics , Iridoids/pharmacology , Fluconazole/pharmacology , Antifungal Agents/pharmacology , Drug Resistance, Fungal , Anti-Bacterial Agents/pharmacology , Microscopy, Electron, Scanning
18.
Front Cell Infect Microbiol ; 14: 1322847, 2024.
Article in English | MEDLINE | ID: mdl-38707513

ABSTRACT

The aetiology of chronic aseptic meningitis is difficult to establish. Candida meningitis in particular is often diagnosed late, as cerebrospinal fluid (CSF) work-up and imaging findings are nonspecific. A 35-year-old patient with chronic aseptic meningitis, for which repeated microbiological testing of CSF was unrevealing, was finally diagnosed with Candida albicans (C. albicans) meningitis with cauda equina involvement using metagenomic next-generation sequencing (mNGS). This report highlights the diagnostic challenges and the difficulties of treating shunt-associated fungal meningitis.


Subject(s)
Candida albicans , High-Throughput Nucleotide Sequencing , Meningitis, Fungal , Metagenomics , Humans , Adult , Candida albicans/genetics , Candida albicans/isolation & purification , Meningitis, Fungal/diagnosis , Meningitis, Fungal/microbiology , Meningitis, Fungal/drug therapy , Metagenomics/methods , Candidiasis/diagnosis , Candidiasis/microbiology , Candidiasis/cerebrospinal fluid , Male , Chronic Disease , Antifungal Agents/therapeutic use , Meningitis, Aseptic/diagnosis
19.
Sci Rep ; 14(1): 10228, 2024 05 03.
Article in English | MEDLINE | ID: mdl-38702418

ABSTRACT

Chitosan (CH) exhibits low antimicrobial activity. This study addresses this issue by modifying the chitosan with a sulfonamide derivative, 3-(4-(N,N-dimethylsulfonyl)phenyl)acrylic acid. The structure of the sulfonamide-chitosan derivative (DMS-CH) was confirmed using Fourier transform infrared spectroscopy and Nuclear magnetic resonance. The results of scanning electron microscopy, thermal gravimetric analysis, and X-ray diffraction indicated that the morphology changed to a porous nature, the thermal stability decreased, and the crystallinity increased in the DMS-CH derivative compared to chitosan, respectively. The degree of substitution was calculated from the elemental analysis data and was found to be moderate (42%). The modified chitosan exhibited enhanced antimicrobial properties at low concentrations, with a minimum inhibitory concentration (MIC) of 50 µg/mL observed for B. subtilis and P. aeruginosa, and a value of 25 µg/mL for S. aureus, E. coli, and C. albicans. In the case of native chitosan, the MIC values doubled or more, with 50 µg/mL recorded for E. coli and C. albicans and 100 µg/mL recorded for B. subtilis, S. aureus, and P. aeruginosa. Furthermore, toxicological examinations conducted on MCF-7 (breast adenocarcinoma) cell lines demonstrated that DMS-CH exhibited greater toxicity (IC50 = 225.47 µg/mL) than pure CH, while still maintaining significant safety limits against normal lung fibroblasts (WI-38). Collectively, these results suggest the potential use of the newly modified chitosan in biomedical applications.


Subject(s)
Anti-Infective Agents , Chitosan , Microbial Sensitivity Tests , Sulfonamides , Chitosan/chemistry , Chitosan/pharmacology , Humans , Sulfonamides/pharmacology , Sulfonamides/chemistry , Anti-Infective Agents/pharmacology , Anti-Infective Agents/chemistry , Candida albicans/drug effects , Staphylococcus aureus/drug effects , Escherichia coli/drug effects , Spectroscopy, Fourier Transform Infrared , Cell Survival/drug effects , X-Ray Diffraction , MCF-7 Cells
20.
Front Cell Infect Microbiol ; 14: 1404960, 2024.
Article in English | MEDLINE | ID: mdl-38803574

ABSTRACT

Staphylococcus aureus and Staphylococcus epidermidis stand as notorious threats to human beings owing to the myriad of infections they cause. The bacteria readily form biofilms that help in withstanding the effects of antibiotics and the immune system. Intending to combat the biofilm formation and reduce the virulence of the pathogens, we investigated the effects of carotenoids, crocetin, and crocin, on four Staphylococcal strains. Crocetin was found to be the most effective as it diminished the biofilm formation of S. aureus ATCC 6538 significantly at 50 µg/mL without exhibiting bactericidal effect (MIC >800 µg/mL) and also inhibited the formation of biofilm by MSSA 25923 and S. epidermidis at a concentration as low as 2 µg/mL, and that by methicillin-resistant S. aureus MW2 at 100 µg/mL. It displayed minimal to no antibiofilm efficacy on the Gram-negative strains Escherichia coli O157:H7 and Pseudomonas aeruginosa as well as a fungal strain of Candida albicans. It could also curb the formation of fibrils, which partly contributes to the biofilm formation in S. epidermidis. Additionally, the ADME analysis of crocetin proclaims how relatively non-toxic the chemical is. Also, crocetin displayed synergistic antibiofilm characteristics in combination with tobramycin. The presence of a polyene chain with carboxylic acid groups at its ends is hypothesized to contribute to the strong antibiofilm characteristics of crocetin. These findings suggest that using apocarotenoids, particularly crocetin might help curb the biofilm formation by S. aureus and S. epidermidis.


Subject(s)
Anti-Bacterial Agents , Biofilms , Carotenoids , Microbial Sensitivity Tests , Staphylococcus epidermidis , Vitamin A , Biofilms/drug effects , Carotenoids/pharmacology , Vitamin A/analogs & derivatives , Vitamin A/pharmacology , Anti-Bacterial Agents/pharmacology , Staphylococcus epidermidis/drug effects , Candida albicans/drug effects , Staphylococcus aureus/drug effects , Humans , Pseudomonas aeruginosa/drug effects , Staphylococcus/drug effects
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