RESUMEN

Introduction. Tissue conditioners modified with antifungals are a potential alternative to denture stomatitis (DS) treatment.Gap Statement. Information on tissue response to this treatment before its clinical application is lacking.Aim. This study aimed to evaluate the tissue response of a tissue conditioner modified with antifungals in a rat model of DS.Methodology. After DS induction for 4 days under antibiotic therapy, Wistar rats had their intraoral devices (IODs) relined with the tissue conditioner Softone without (Soft) or with the MICs against Candida albicans of nystatin (Nys) or chlorhexidine (Chx) complexed or not with ß-cyclodextrin (Nys:ßCD and Chx:ßCD). Three controls were included: healthy rats [negative control (Nc)], rats using a sterile IOD [sterile device (Sd)] and rats with DS that did not receive treatment (DS). After 4 days of treatment, the palatal mucosa under the IODs underwent histological processing for morphohistopathological and histometric analyses, morphology of collagen fibres (birefringence), immunohistochemistry for the expression of cell proliferation (proliferating cell nuclear antigen) and cytokine (IL-1ß).Results. The Nc and Sd groups were similar (P>0.05), displaying epithelial and connective tissues without any discernible changes in the parameters assessed. The DS and Soft groups exhibited pronounced epithelial alterations, cell proliferation and expression of the cytokine IL-1ß. In groups treated with drug incorporation (Nys, Chx, Nys:ßCD and Chx:ßCD), all samples demonstrated a reduction in tissue inflammation or complete tissue recovery, with an epithelium compatible with health. For the immunohistochemical parameters, the Chx, Nys:ßCD and Chx:ßCD groups were comparable with Nc (P>0.05).Conclusion. The proposed treatment could be promising for DS, as it led to the tissue recovery of the palatal mucosa. Nevertheless, much lower concentrations of complexed antifungals were required to achieve a similar or higher degree of tissue response compared with uncomplexed drugs in a modified tissue conditioner formulation.

Asunto(s)

Antifúngicos , Candida albicans , Modelos Animales de Enfermedad , Mucosa Bucal , Nistatina , Ratas Wistar , Estomatitis Subprotética , beta-Ciclodextrinas , Animales , beta-Ciclodextrinas/química , Antifúngicos/farmacología , Estomatitis Subprotética/tratamiento farmacológico , Estomatitis Subprotética/microbiología , Ratas , Nistatina/farmacología , Nistatina/administración & dosificación , Candida albicans/efectos de los fármacos , Mucosa Bucal/efectos de los fármacos , Mucosa Bucal/microbiología , Masculino , Clorhexidina/farmacología , Interleucina-1beta/metabolismo , Antígeno Nuclear de Célula en Proliferación/metabolismo , Pruebas de Sensibilidad Microbiana

RESUMEN

This data descriptor presents a curated dataset for pathogen detection and identification (Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans) directly from whole-blood samples. The dataset was created using differential cell lysis combined with rapid extraction, digestion, and mass spectrometry-based proteomics. Our method offers a rapid diagnostic alternative to traditional culture, enabling timely disease management, such as sepsis. Highlighting our dataset's uniqueness, it features a three-tier structure: Spectral Libraries of Pathogens for identifying peptide peaks for putative biomarkers; Spiked pathogen in blood MS data for biomarker panel optimization through varied concentration samples; and Parallel Reaction Monitoring (PRM) data from sepsis patients for validating our biomarker panel, achieving 83.3% sensitivity within seven hours without microbial enrichment culture. This dataset serves as a comprehensive reference for bioinformatic tool development and biomarker panel proposals, advancing microbial detection, antimicrobial resistance, and epidemiological studies.

Asunto(s)

Biomarcadores , Candida albicans , Proteómica , Pseudomonas aeruginosa , Sepsis , Staphylococcus aureus , Humanos , Proteómica/métodos , Biomarcadores/sangre , Sepsis/sangre , Sepsis/diagnóstico , Sepsis/microbiología , Espectrometría de Masas

RESUMEN

Candida albicans is often associated with oral candidiasis, and drug-resistance profiles have contributed to an increase in morbidity and mortality. It is known that Lactobacillus spp. acts by competing for adhesion to the epithelium, absorption of nutrients and modulation of the human microbiota. Therefore, they are important to assist in the host's microbiological balance and reduce the growth of Candida spp. Until now, there have been no reports in the literature of reviews correlating to the use of Lactobacillus spp. in the treatment of oral candidiasis. Thus, this review aims to highlight the mechanisms of action of Lactobacillus spp. and methods that can be used in the treatment of oral candidiasis. This is a study carried out through the databases PubMed Central and Scientific Electronic Library Online, using the following keywords: Oral Candidiasis and Lactobacillus. Original articles about oral candidiasis were included, with both in vitro and in vivo analyses, and published from 2012 to 2022. Lactobacillus rhamnosus was the most common microorganism used in the experiments against Candida, acting mainly in the reduction of biofilm, filamentation, and competing for adhesion sites of Candida spp. Among in vivo studies, most researchers used immunosuppressed mouse modelsof Candida infection. The studies showed that Lactobacillus has a great potential as a probiotic, acting mainly in the prevention and treatment of mucosal diseases. Thus, the use of Lactobacillus may be a good strategy for the treatment of oral candidiasis.

Asunto(s)

Candidiasis Bucal , Lactobacillus , Probióticos , Candidiasis Bucal/microbiología , Candidiasis Bucal/terapia , Candidiasis Bucal/tratamiento farmacológico , Humanos , Probióticos/uso terapéutico , Lactobacillus/fisiología , Animales , Biopelículas/efectos de los fármacos , Ratones , Candida albicans/fisiología

RESUMEN

Candida albicans invasive candidiasis is considered a global health problem. In such cases, biofilm formation on implanted devices represents a therapeutic challenge and the presence of metabolically inactive persistent cells (PCs) in these communities increases their tolerance to fungicidal drugs. This study investigated the influence of amoxicillin, AMX; cefepime, CEF; gentamicin, GEN; amikacin, AMK; vancomycin, VAN; and ciprofloxacin, CIP; on the production of PCs in biofilms of C. albicans bloodstream isolates. 48 h-mature biofilms (n = 6) grown in RPMI-1640 supplemented with antibiotics were treated with 100 µg ml-1 amphotericin B and then evaluated for PCs. Biofilms grown in the presence of antibiotics produced more PCs, up to 10×, when exposed to AMX and CIP; 5 × to CEF; and 6 × to GEN and VAN. The results indicate that antibiotics can modulate PC production in C. albicans biofilms. This scenario may have clinical repercussions in immunocompromised patients under broad-spectrum antibiotic therapy.

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Biofilms are microbial communities tolerant to antifungals. Our research showed that antibiotics stimulate the formation of persistent cells within Candida albicans biofilms. These are dormant, metabolically silent cells that resist to therapy and can be related to metastatic and recalcitrant infections.

Asunto(s)

Antibacterianos , Biopelículas , Candida albicans , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Candida albicans/efectos de los fármacos , Candida albicans/fisiología , Antibacterianos/farmacología , Humanos , Antifúngicos/farmacología , Pruebas de Sensibilidad Microbiana , Ciprofloxacina/farmacología , Gentamicinas/farmacología , Amoxicilina/farmacología , Vancomicina/farmacología , Amicacina/farmacología , Cefepima/farmacología , Anfotericina B/farmacología , Cefalosporinas/farmacología , Candidiasis/microbiología , Candidiasis/tratamiento farmacológico

RESUMEN

The primary aim of this study was to investigate the impact of treatment with low-temperature plasma (LTP) for varying exposure durations on a multispecies cariogenic biofilm comprising C. albicans, L. casei, and S. mutans, as well as on single-species biofilms of L. casei and C. albicans, cultured on hydroxyapatite discs. Biofilms were treated with LTP-argon at a 10 mm distance for 30 s, 60 s, and 120 s. Chlorhexidine solution (0.12%) and NaCl (0.89%) were used as positive (PC) and negative controls (NC), respectively. Argon flow only was also used as gas flow control (F). Colony-forming units (CFU) recovery and confocal laser scanning microscopy (CLSM) were used to analyze biofilm viability. LTP starting at 30 s of application significantly reduced the viability of multispecies biofilms by more than 2 log10 in all treated samples (p < 0.0001). For single-species biofilms, L. casei showed a significant reduction compared to PC and NC of over 1 log10 at all exposure times (p < 0.0001). In the case of C. albicans biofilms, LTP treatment compared to PC and NC resulted in a significant decrease in bacterial counts when applied for 60 and 120 s (1.55 and 1.90 log10 CFU/mL, respectively) (p < 0.0001). A significant effect (p ≤ 0.05) of LTP in single-species biofilms was observed to start at 60 s of LTP application compared to F, suggesting a time-dependent effect of LTP for the single-species biofilms of C. albicans and L. casei. LTP is a potential mechanism in treating dental caries by being an effective anti-biofilm therapy of both single and multispecies cariogenic biofilms.

Asunto(s)

Biopelículas , Candida albicans , Gases em Plasma , Streptococcus mutans , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Gases em Plasma/farmacología , Candida albicans/fisiología , Candida albicans/efectos de los fármacos , Streptococcus mutans/efectos de los fármacos , Streptococcus mutans/fisiología , Caries Dental/microbiología , Caries Dental/terapia , Lacticaseibacillus casei/fisiología , Humanos , Viabilidad Microbiana/efectos de los fármacos , Microscopía Confocal , Frío

RESUMEN

This study proposes an affordable plasma device that utilizes a parallel-plate dielectric barrier discharge geometry with a metallic mesh electrode, featuring a straightforward 3D-printed design. Powered by a high-voltage supply adapted from a cosmetic plasma device, it operates on atmospheric air, eliminating the need for gas flux. Surface modification of polyethylene treated with this device was characterized and showed that the elemental composition after 15 min of plasma treatment decreased the amount of C to ~80 at% due to the insertion of O (~15 at%). Tested against Candida albicans and Staphylococcus aureus, the device achieved a reduction of over 99% in microbial load with exposure times ranging from 1 to 10 min. Simultaneously, the Vero cell viability remained consistently high, namely between 91% and 96% across exposure times. These results highlight this device's potential for the surface modification of materials and various infection-related applications, boasting affordability and facilitating effective antimicrobial interventions.

Asunto(s)

Candida albicans , Gases em Plasma , Staphylococcus aureus , Propiedades de Superficie , Candida albicans/efectos de los fármacos , Gases em Plasma/química , Gases em Plasma/farmacología , Staphylococcus aureus/efectos de los fármacos , Animales , Células Vero , Chlorocebus aethiops , Viabilidad Microbiana/efectos de los fármacos , Polímeros/química

RESUMEN

The immunopathogenesis of recurrent vulvovaginal candidiasis (RVVC) is poorly understood. Recently, it was reported that patients with RVVC present a decrease in both the fungicidal capacity of neutrophils and the proliferative capability of peripheral blood mononuclear cells in response to Candida albicans infection, suggesting an alteration in the innate and adaptive immune response. The aim of this study was to determine the in-situ expression, in the vaginal mucosa, of genes associated with the immune response, as well as the serum concentrations of dectin-1, mannose-binding lectin (MBL), and vitamin D in patients with RVVC. A study was carried out on 40 patients with a diagnosis of RVVC and 26 healthy women. Vaginal scrapings were obtained, and the expression of genes that encode cytokines and transcription factors specific for Th1, Th2, Th17, Treg, pro-inflammatory profiles, and enzymes related to oxidative/microbicidal mechanisms was evaluated by quantitiative polymerase chain reaction (qPCR). Additionally, serum levels of vitamin D and the soluble receptors dectin-1 and MBL were determined by enzyme-linked immunosorbent assay (ELISA). In patients with RVVC, a decreased expression of T-bet, RORγ-T, IL-1ß, and IL-17, and an increase in the expression of FOXP3, IL-4, IL-8, IL-10, and IL-18 were observed when compared to healthy women: moreover, decreased levels of MBL were also observed in these patients. These results confirm that patients with RVVC present in-situ alterations in both the specific and adaptive immune response against Candida spp., a fact that could be associated with the exaggerated vaginal inflammatory response.

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The study concerns the immune response of women with recurrent vulvovaginal candidiasis; we observed an alteration in the expression of genes that participate in the control of infection, a fact that could be associated with the exaggerated vaginal inflammatory response observed in those patients.

Asunto(s)

Candidiasis Vulvovaginal , Citocinas , Lectinas Tipo C , Vagina , Vitamina D , Humanos , Candidiasis Vulvovaginal/inmunología , Candidiasis Vulvovaginal/microbiología , Femenino , Lectinas Tipo C/genética , Adulto , Citocinas/sangre , Vagina/microbiología , Vagina/inmunología , Vitamina D/sangre , Adulto Joven , Recurrencia , Lectina de Unión a Manosa/sangre , Lectina de Unión a Manosa/genética , Membrana Mucosa/inmunología , Membrana Mucosa/microbiología , Candida albicans/inmunología

RESUMEN

The discovery that the lung harbors a diverse microbiome, as revealed by next-generation sequencing, has significantly altered our understanding of respiratory health and disease. Despite the association between the lung microbiota and disease, the nature of their relationship remains poorly understood, and culture isolation of these microorganisms could help to determine their role in lung physiology. Current procedures for processing samples from the lower respiratory tract have been shown to affect the viability of microorganisms, so it is crucial to develop new methods to improve their survival. This study aimed to improve the isolation and characterization of lung microorganisms using a bead-beating homogenization method in a mouse model. Microsphere diameter and bead-beating time affected the survival of the microorganisms (E. coli, S. aureus and C. albicans). Using 2.3 mm diameter microspheres for 60 s of bead-beating promoted the survival of both bacteria and yeast strains. After intratracheal instillation of these microorganisms in mice, approximately 70% of the cells were recovered after the tissue homogenization. To assess the efficiency of the proposed method, the diversity of bacteria was compared between the homogenate and lung tissue samples. Ninety-one genera were detected in the lung tissue, and 63 in the homogenate. Bacterial genera detected in the homogenate represented 84% of the total abundance of the microbiota identified in the lung tissue. Taken together, these results demonstrate that the tissue homogenization process developed in this study recovered the majority of the microorganisms present in the lung. This study presents a bead-beating homogenization method for effective cultivation of lung tissue microorganisms, which may help to improve the understanding of host-microbe interactions in the lung.

Asunto(s)

Pulmón , Microbiota , Animales , Pulmón/microbiología , Ratones , Microesferas , Staphylococcus aureus , Candida albicans/aislamiento & purificación , Escherichia coli/aislamiento & purificación , Bacterias/aislamiento & purificación , Bacterias/clasificación , Bacterias/genética

RESUMEN

Aim: Synthetic antimicrobial peptides (SAMPs) present the potential to fight systemic fungal infections. Here, the PHO36 receptor from Candida albicans was analyzed by in silico tools as a possible target for three anticandidal SAMPs: RcAlb-PepIII, PepGAT and PepKAA.Materials & methods: Molecular docking, dynamics and quantum biochemistry were employed to understand the individual contribution of amino acid residues in the interaction region.Results: The results revealed that SAMPs strongly interact with the PHO36 by multiple high-energy interactions. This is the first study to employ quantum biochemistry to describe the interactions between SAMPs and the PHO36 receptor.Conclusion: This work contributes to understanding and identifying new molecular targets with medical importance that could be used to discover new drugs against systemic fungal infections.

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Here, computers helped us find new proteins in Candida albicans that may guide the development of new medicines.

Asunto(s)

Antifúngicos , Candida albicans , Simulación del Acoplamiento Molecular , Candida albicans/efectos de los fármacos , Antifúngicos/farmacología , Antifúngicos/química , Antifúngicos/síntesis química , Péptidos Antimicrobianos/farmacología , Péptidos Antimicrobianos/química , Péptidos Antimicrobianos/síntesis química , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/genética , Simulación de Dinámica Molecular , Simulación por Computador , Unión Proteica , Humanos

RESUMEN

Patients with orofacial clefts are more likely to develop oral fungal diseases due to anatomo-physiological changes and surgical rehabilitation treatment. This case-series study evaluated the genetic diversity and dynamics of oral colonization and spread of C. albicans and C. tropicalis in four patients with orofacial clefts, from the time of hospital admission, perioperative and outpatient follow-up, with specialized physician. Candida biotypes previously identified by CHROMagar Candida and PCR methods were studied by MALDI-TOF MS assays and clustering analyses. Possible correlations with pathogenicity characteristics were observed, including production of hydrolytic exoenzymes and the antifungal sensitivity profiles. Amphotericin B-sensitive and fluconazole-resistant (low frequency) C. tropicalis and C. albicans, including clinically compatible MIC of nystatin, were found in the oral cavity of these patients. Clusters of isolates revealed phenomena of (i) elimination in the operative phase, (ii) maintenance or (iii) acquisition of oral C. tropicalis in the perioperative period and specialized outpatient and medical follow-up. For C. albicans, these phenomena included (i) elimination in the operative phase, (ii) acquisition in the operative phase and propagation from the hospital environment, and (iii) maintenance during hospitalization and operative phase. Amphotericin B and nystatin were shown to be effective in cases of clinical treatment and/or prophylaxis, especially considering the pre-existence of fluconazole-resistant strains. This study confirmed the phenomena of septic maintenance, septic neocolonization and septic elimination involving the opportunistic pathogens. MALDI-TOF MS associated with clustering analysis may assist the monitoring of clinical isolates or groups of epidemiologically important microbial strains in the hospital setting.

Asunto(s)

Anfotericina B , Antifúngicos , Candida albicans , Candida tropicalis , Farmacorresistencia Fúngica , Genotipo , Pruebas de Sensibilidad Microbiana , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Humanos , Candida tropicalis/efectos de los fármacos , Candida tropicalis/aislamiento & purificación , Candida tropicalis/genética , Antifúngicos/farmacología , Antifúngicos/uso terapéutico , Candida albicans/genética , Candida albicans/efectos de los fármacos , Candida albicans/aislamiento & purificación , Anfotericina B/farmacología , Anfotericina B/uso terapéutico , Masculino , Femenino , Boca/microbiología , Niño , Fluconazol/farmacología , Fluconazol/uso terapéutico , Candidiasis Bucal/microbiología , Nistatina/farmacología , Nistatina/uso terapéutico , Técnicas de Tipificación Micológica , Fisura del Paladar/cirugía , Labio Leporino/cirugía , Adolescente , Análisis por Conglomerados , Preescolar

RESUMEN

INTRODUCTION: Candida albicans is the most common opportunistic pathogen causing fungal infections worldwide, especially in high-risk patients. Its pathogenicity is related to virulence factors gene expression, such as hyphal growth (HWP1), cell adhesion (ALS3), and protease secretion (SAP1) during infection spreading mechanisms. In recent years, an increase in non-albicans Candida infections has been reported, which may present coinfection or competitive interactions with C. albicans, potentially aggravating the patient's condition. This study aims to evaluate the expression of genes related to virulence factors of C. albicans and non-albicans Candida during planktonic stage. METHODS: C. albicans (ATCC MYA-3573) as well as with three clinical strains (C. albicans DCA53, C. tropicalis DCT6, and C. parapsilosis DCP1) isolated from blood samples, were grown in 24-well plates at 37°C for 20 h, either in monocultures or mixed cultures. Quantitative real-time polymerase chain reaction was used to evaluate the expression levels of the genes HWP1, ALS3, and SAP1 in cells collected during the planktonic stage. In addition, hyphal filamentation was observed using a Scanning Electron Microscope. RESULTS: The overexpression of HWP1 and ASL3 genes in mixed growth conditions between C. albicans and non-albicans Candida species suggests a synergistic relationship as well as an increased capacity for hyphal growth and adhesion. In contrast, C. parapsilosis versus C. tropicalis interaction shows an antagonistic relationship during mixed culture, suggesting a decreased virulence profile of C. parapsilosis during initial coinfection with C. tropicalis. CONCLUSION: The expression of HWP1, ALS3, and SAP1 genes associated with virulence factors varies under competitive conditions among species of the genus Candida during planktonic stage.

Asunto(s)

Candida albicans , Proteínas Fúngicas , Factores de Virulencia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Humanos , Factores de Virulencia/genética , Candida albicans/patogenicidad , Candida albicans/genética , Virulencia/genética , Hifa/genética , Regulación Fúngica de la Expresión Génica , Candidiasis/microbiología , Ácido Aspártico Endopeptidasas/genética , Ácido Aspártico Endopeptidasas/metabolismo , Plancton/genética , Candida/patogenicidad , Candida/genética , Glicoproteínas de Membrana

RESUMEN

OBJECTIVE: Adhesion to dentin is a first step for a successful microbial root canal colonization. Cell hydrophobicity seems to have some influence in the Candida species adhesion to surfaces. To measure cell surface hydrophobicity and to investigate the adherence ability to human dentin among Candida albicans strains isolated from root canal and lingual dorsum via an in vitro study. METHODOLOGY: adhesion was quantified in function of dentin area covered by blastospores and/or hyphae presence detected by epifluorescence microscope. Cell surface hydrophobicity was estimated by assessing the percentage migration of cells from an aqueous phase to a hydrocarbon phase. Contact angles were measured by the sessile drop technique on the dentin surface using a contact angle measurements apparatus. We also examined the correlation between adhesion ability and hydrophobicity. RESULTS: although there was some intra-species variation in cell surface hydrophobicity, most isolates were characterized by moderate hydrophobicity. There was no significant difference in this parameter when the isolation niche was considered. Both root canal and lingual dorsum yeasts were able to adhere to dentin. No association was found between the strains' site of isolation and adhesion. Moreover, cell surface hydrophobicity and adhesion ability were not correlated. CONCLUSION: although hydrophobicity can influence Candida albicans virulence in many ways, this study suggests that this parameter by itself was not a good predictor of adhesion to dentin.

Asunto(s)

Candida albicans , Adhesión Celular , Dentina , Interacciones Hidrofóbicas e Hidrofílicas , Propiedades de Superficie , Candida albicans/fisiología , Humanos , Dentina/microbiología , Adhesión Celular/fisiología , Cavidad Pulpar/microbiología , Microscopía Fluorescente

RESUMEN

Both Candida albicans and Issatchenkia orientalis have been isolated from different types of infections over the years. They have the ability to form communities of microorganisms known as biofilms. It has been demonstrated that the medium employed in studies may affect the biofilm development. The aim of this study was to investigate the arrangement of dual-species biofilms of C. albicans and I. orientalis cultivated on either RPMI-1640 or Sabouraud Dextrose Broth (SDB), as well as the inhibitory effect of Voriconazole (VRC). For the experiments performed, ATCC strains were used, and yeast-mixed suspensions were inoculated in 96-well plates with either RPMI-1640 or SDB, in the presence or absence of VRC. The results were observed by counting the number of CFU obtained from scraping off the biofilms produced and plating the content on CHROMagar Candida medium. It was observed that for all conditions tested the medium chosen affected the arrangement of dual-species biofilms: when RPMI-1640 was used, there was a prevalence of C. albicans, while the opposite was noted when SDB was used. It could be suggested that the medium and environment could regulate interactions between both yeast species, including the response to different antifungal drugs.

Asunto(s)

Antifúngicos , Biopelículas , Candida albicans , Medios de Cultivo , Voriconazol , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Voriconazol/farmacología , Candida albicans/efectos de los fármacos , Candida albicans/fisiología , Antifúngicos/farmacología , Medios de Cultivo/química , Saccharomycetales/efectos de los fármacos , Saccharomycetales/fisiología , Pruebas de Sensibilidad Microbiana

RESUMEN

Plant peptidase inhibitors play crucial roles in plant defence mechanisms and physiological processes. In this study, we isolated and characterised a Kunitz trypsin inhibitor from Enterolobium gummiferum seeds named EgPI (E. gummiferum peptidase inhibitor). The purification process involved two chromatography steps using size exclusion and hydrophobic resins, resulting in high purity and yield. EgPI appeared as a single band of ~20 kDa in SDS-PAGE. Under reducing conditions, the inhibitor exhibited two polypeptide chains, with 15 and 5 kDa. Functional characterisation revealed that EgPI displayed an inhibition stoichiometry of 1:1 against trypsin, with a dissociation constant of 8.4 × 10-9 mol·L-1. The amino-terminal sequencing of EgPI revealed the homology with Kunitz inhibitors. Circular dichroism analysis provided insights into the secondary structure of EgPI, which displayed the signature typical of Kunitz inhibitors. Stability studies demonstrated that EgPI maintained the secondary structure necessary to exhibit its inhibitory activity up to 70 °C and over a pH range from 2 to 8. Microbiological screening revealed that EgPI has antibiofilm properties against pathogenic yeasts at 1.125 µmol·L-1, and EgPI reduced C. albicans biofilm formation by 82.7%. The high affinity of EgPI for trypsin suggests potential applications in various fields. Furthermore, its antibiofilm properties recommended its usefulness in agriculture and antimicrobial therapy research, highlighting the practical implications of our research.

Asunto(s)

Biopelículas , Fabaceae , Proteínas de Plantas , Semillas , Inhibidores de Tripsina , Semillas/química , Biopelículas/efectos de los fármacos , Fabaceae/química , Inhibidores de Tripsina/farmacología , Inhibidores de Tripsina/química , Inhibidores de Tripsina/aislamiento & purificación , Proteínas de Plantas/farmacología , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificación , Candida albicans/efectos de los fármacos , Antifúngicos/farmacología , Antifúngicos/química , Antifúngicos/aislamiento & purificación , Secuencia de Aminoácidos , Péptidos

RESUMEN

Vulvovaginal candidiasis (VVC) alters the innate cervicovaginal immunity, which provides an important barrier against viruses and other infections. The incidence of this disease has not decreased in the last 30 years, so effective treatments are still needed. Nanoparticles (NPs) of cellulose acetate phthalate (CAP) and clotrimazole (CLZ) were prepared by the emulsification-diffusion method. NPs were characterized using dynamic light scattering, atomic force microscopy and differential scanning calorimetry; their release profile was determined by the dialysis bag technique and mucoadhesion was evaluated with the mucin-particle method. The growth inhibition study of Candida albicans was carried out using the plate counting technique. Finally, accelerated physical stability tests of NPs were carried out, both in water and in SVF. The CAP-CLZ NPs had an average diameter of 273.4 nm, a PDI of 0.284, smooth surfaces and spherical shapes. In vitro release of CLZ from the CAP NPs was categorized with the Weibull model as a matrix system in which initial release was rapid and subsequently sustained. The inhibition of C. albicans growth by the CAP-CLZ NPs was greater than that of free CLZ, and the CAP-only NPs had a microbicidal effect on C. albicans. The NPs showed poor mucoadhesiveness, which could lead to studies of their mucopenetration capacities. An accelerated physical stability test revealed the erosion of CAP in aqueous media. A nanoparticulate system was developed and provided sustained release of CLZ, and it combined an antifungal agent with a microbial polymer that exhibited antifungal activity against C. albicans.

Asunto(s)

Antifúngicos , Candida albicans , Candidiasis Vulvovaginal , Celulosa , Clotrimazol , Nanopartículas , Clotrimazol/administración & dosificación , Clotrimazol/farmacología , Candidiasis Vulvovaginal/tratamiento farmacológico , Nanopartículas/química , Candida albicans/efectos de los fármacos , Femenino , Celulosa/química , Celulosa/análogos & derivados , Antifúngicos/farmacología , Antifúngicos/administración & dosificación , Polímeros/química , Tamaño de la Partícula , Pruebas de Sensibilidad Microbiana/métodos , Liberación de Fármacos

RESUMEN

This study investigated crotamine (CTA), a peptide derived from the venom of the South American rattlesnake Crotalus durissus terrificus, known for its exceptional cell penetration potential. The objective was to explore the antibacterial and antifungal activity of CTA, its ability to inhibit efflux pumps and evaluate the effectiveness of its pharmacological combination with antibiotics and antifungals. In microbiological assays, CTA in combination with antibiotics was tested against strains of S. aureus and the inhibition of NorA, Tet(K) and MepA efflux pumps was also evaluated. CTA alone did not present clinically relevant direct antibacterial action, presenting MIC > 209.7 µM against strains S. aureus 1199B, IS-58, K2068. The standard efflux pump inhibitor CCCP showed significant effects in all negative relationships to assay reproducibility. Against the S. aureus 1199B strain, CTA (20.5 µM) associated with norfloxacin diluted 10 × (320.67 µM) showed a potentiating effect, in relation to the control. Against the S. aureus IS-58 strain, the CTA associated with tetracycline did not show a significant combinatorial effect, either with 2304 or 230.4 µM tetracycline. CTA at a concentration of 2.05 µM associated with ciprofloxacin at a concentration of 309.4 µM showed a significant potentiating effect. In association with EtBr, CTA at concentrations of 2.05 and 20.5 µM potentiated the effect in all strains tested, reducing the prevention of NorA, Tet(K) and MepA efflux pumps. In the C. albicans strain, a potentiating effect of fluconazole (334.3 µM) was observed when combined with CTA (2.05 µM). Against the C. tropicalis strain, a significant effect was also observed in the association of fluconazole 334.3 µM, where CTA 2.05 µM considerably reduced fungal growth and decreased the potentiation of fluconazole. Against the C. krusei strain, no significant potentiating effect of fluconazole was obtained by CTA. Our results indicate that CTA in pharmacological combination potentiates the effects of antibiotics and antifungal. This represents a new and promising antimicrobial strategy for treating a wide variety of infections.

Asunto(s)

Antibacterianos , Antifúngicos , Venenos de Crotálidos , Crotalus , Pruebas de Sensibilidad Microbiana , Antifúngicos/farmacología , Antifúngicos/química , Antibacterianos/farmacología , Venenos de Crotálidos/farmacología , Animales , Staphylococcus aureus/efectos de los fármacos , Sinergismo Farmacológico , Candida albicans/efectos de los fármacos , Serpientes Venenosas

RESUMEN

Because of the exoskeleton, arthropods must have flexible areas to be able to move. Such regions are called arthrodial membranes and are particularly vulnerable to bacteria and fungi. Here, we analyzed the secretion in the glands underneath it in a Neotropical harvester Mischonyx squalidus (Arachnida: Opiliones) and tested whether it has antiseptical properties. Wepuncturedthemembrane,collectedand quantified ina spectrophotometer. We also fractionated and analyzed the samples in reversed-phase high-performance liquid chromatography (RP-HPLC) and then incubated the treated fractions and determined growth inhibition by measuring absorbance. The secretions resulted in 100 fractions, among which two had activity against the Gram-positive bacteria Micrococcus luteus and against the yeast Candida albicans. The low concentrations at which the secretions were active are relevant from a biotechnological point of view. For the organism, the secretions possibly prevent infections, including when they are attacked in these regions by predators that pick that spot to bite.

Asunto(s)

Arácnidos , Animales , Arácnidos/metabolismo , Candida albicans/efectos de los fármacos , Micrococcus luteus/efectos de los fármacos , Antiinfecciosos/metabolismo , Antiinfecciosos/química , Antiinfecciosos/farmacología

RESUMEN

The use of photothermal processes has been proven effective in the control of microbial infections. Simultaneously, the localized surface plasmon resonance phenomena in metallic nanoparticles have been explored as an alternative strategy to achieve highly efficient localized heating. In this work, we propose the use of selected nanoheaters to improve the efficiency of fungal photothermal inactivation of Candida albicans through size optimization of plasmonic gold nanorods. Here, the optical heating of polyethylene glycol coated gold nanorods of varying sizes is evaluated, both theoretically and experimentally. A size-dependent computational approach was applied to identify metallic nanorods with maximized thermal performance at 800 nm, followed by the experimental comparison of optimal and suboptimal nanoheaters. Comparison among samples show temperatures of up to 53.0 °C for 41×10 nm gold nanorods against 32.3 °C for 90×25 nm, a percentage increase of ∼63% in photothermal inactivation assessments. Our findings reveal that gold nanorods of 41×10 nm exhibit superior efficiency in near-infrared (800 nm) photothermal inactivation of fungi, owing to their higher light-thermal conversion efficiency. The identification of high performance metallic nanoheaters may lead to the reduction of the nanoparticle dose used in plasmonic-based procedures and decrease the laser exposure time needed to induce cell death. Moreover, our results provide insights to better exploit plasmonic nanoparticles on photothermal inactivation protocols.

Asunto(s)

Candida albicans , Oro , Nanopartículas del Metal , Nanotubos , Candida albicans/efectos de los fármacos , Nanotubos/química , Oro/química , Oro/farmacología , Nanopartículas del Metal/química , Resonancia por Plasmón de Superficie , Rayos Infrarrojos , Polietilenglicoles/química , Polietilenglicoles/farmacología , Fotoquimioterapia/métodos , Terapia Fototérmica/métodos

RESUMEN

Previous reports have demonstrated that the peptide derived from LfcinB, R-1-R, exhibits anti-Candida activity, which is enhanced when combined with an extract from the Bidens pilosa plant. However, the mechanism of action remains unexplored. In this research, a proteomic study was carried out, followed by a bioinformatic analysis and biological assays in both the SC5314 strain and a fluconazole-resistant isolate of Candida albicans after incubation with R-1-R. The proteomic data revealed that treatment with R-1-R led to the up-regulation of most differentially expressed proteins compared to the controls in both strains. These proteins are primarily involved in membrane and cell wall biosynthesis, membrane transport, oxidative stress response, the mitochondrial respiratory chain, and DNA damage response. Additionally, proteomic analysis of the C. albicans parental strain SC5314 treated with R-1-R combined with an ethanolic extract of B. pilosa was performed. The differentially expressed proteins following this combined treatment were involved in similar functional processes as those treated with the R-1-R peptide alone but were mostly down-regulated (data are available through ProteomeXchange with identifier PXD053558). Biological assays validated the proteomic results, evidencing cell surface damage, reactive oxygen species generation, and decreased mitochondrial membrane potential. These findings provide insights into the complex antifungal mechanisms of the R-1-R peptide and its combination with the B. pilosa extract, potentially informing future studies on natural product derivatives.

Asunto(s)

Antifúngicos , Bidens , Candida albicans , Extractos Vegetales , Proteómica , Antifúngicos/farmacología , Proteómica/métodos , Bidens/química , Extractos Vegetales/farmacología , Extractos Vegetales/química , Candida albicans/efectos de los fármacos , Sinergismo Farmacológico , Proteínas Fúngicas/metabolismo , Péptidos/farmacología , Péptidos/química , Pruebas de Sensibilidad Microbiana , Farmacorresistencia Fúngica/efectos de los fármacos , Fluconazol/farmacología

RESUMEN

Candida albicans is the most common agent in human fungal infections; nevertheless, in the last decades, the closely related yeasts Candida dubliniensis and Candida africana have emerged as pathogens. The purpose of this study was to compare tobacco agar with another five agars prepared from plant extracts (Origanum vulgare, Rosmarinus officinalis, Solanum rudepannum, Solanum oblongifolium and Brugmansia arborea) on the differentiation of C. albicans complex. The hyphae and chlamyconidia formation and the color and margin of the colonies of 200 clinical isolates of C. albicans, C. dubliniensis and C. africana were evaluated. After seven days of incubation at 28 °C, Tobacco agar, S. rudepannum and B. arborea agars allowed the differentiation of 100 % C. dubliniensis. Additionally, 24% of C. africana isolates produced brownish colonies in the medium prepared from Rosmarinus officinalis (rosemary) extract. These results indicate that S. rudepannun, B. arborea and rosemary agar could be used as screening for the phenotypic differentiation between the species of C. albicans complex. Rosemary agar could be used to aid in the differentiation of C. albicans from C. africana. These culture media based on plants, could be used as simple and inexpensive screening methods in the phenotypic differentiation of C. dubliniensis and C. africana.

Asunto(s)

Candida albicans , Medios de Cultivo , Extractos Vegetales , Candida albicans/aislamiento & purificación , Candida/clasificación , Candida/aislamiento & purificación , Agar , Humanos , Hifa