Farnesol as a potentiator of antimicrobial photodynamic inactivation on Enterococcus faecalis.

Enterococcus faecalis is related to the recurrence of endodontic infections and approaches to intracanal disinfection are necessary. Farnesol, an alcohol commonly found in propolis, has antimicrobial properties, and can enhance the efficacy of some antibiotic therapies. The objective was to evaluate whether farnesol can increase the efficacy of the antimicrobial photodynamic inactivation (aPDI) on E. faecalis, investigating its action on planktonic growth, biofilms, and cell permeability. Planktonic cells and biofilms of E. faecalis were pre-treated with farnesol (0.25 mM) 2 h before aPDI. Methylene blue (1 mg/mL) and laser (660 nm) were employed in the aPDI. As a result, farnesol was able to increase the antimicrobial activity of aPDI in both planktonic and biofilm stages, reaching cell reductions of 4.6 to 6 log10 CFU and 1.3 to 3 log10 CFU, respectively, when compared to aPDI isolated. The efficacy of farnesol in enhancing the anti-biofilm activity of aPDI was also confirmed by electron microscopy, in which a smaller number of bacterial cells and extracellular matrix were verified in the combined therapy compared to aPDI alone. The potentiating action of farnesol was associated with its effects in increasing the cell permeability and methylene blue uptake by the bacterial cells. Therefore, farnesol can be a promising potentiator of aPDI against E. faecalis.

Enhancing effect of chitosan on methylene blue-mediated photodynamic therapy against C. albicans: A study in planktonic growth, biofilms, and persister cells.

Chitosan (CS) is a natural polymer extracted from the exoskeleton of crustaceans. Due to its cationic structure, CS has been studied as a possible enhancer of antimicrobial photodynamic therapy (aPDT). The objective was to evaluate the association of CS with methylene blue (MB)-mediated aPDT on Candida albicans, investigating its effects on planktonic growth, biofilms, and cells persistent to fluconazole. The ability of CS to interfere with MB absorption by Candida cells was also evaluated. For the assays, planktonic cells of C. albicans were cultivated for 24 h, and the biofilms were formed for 48 h. For the induction of persister cells, C. albicans was cultivated with high concentration of fluconazole for 48 h. Treatments were performed with MB, CS or MB+CS, followed by irradiation with LED (660 nm ). As results, aPDT with MB (300 µm) reduced the planktonic cells by 1.6 log10 CFU, while the MB+CS association led to a reduction of 4.8 log10 CFU. For aPDT in biofilms, there was a microbial reduction of 2.9 log10 CFU for the treatment with MB (600 µm) and 5.3 log10 CFU for MB+CS. In relation to persister cells, the fungal reductions were 0.4 log10 CFU for MB and 1.5 log10 CFU for MB+CS. In the absorption assays, the penetration of MB into Candida cells was increased in the presence of CS. It was concluded that CS enhanced the antimicrobial activity of aPDT in planktonic growth, biofilms, and persister cells of C. albicans, probably by facilitating the penetration of MB into fungal cells.

Cryptococcus spp. and Cryptococcosis: focusing on the infection in Brazil.

Cryptococcosis is a global fungal infection caused by the Cryptococcus neoformans/Cryptococcus gattii yeast complex. This infection is acquired by inhalation of propagules such as basidiospores or dry yeast, initially causing lung infections with the possibility of progressing to the meninges. This infection mainly affects immunocompromised HIV and transplant patients; however, immunocompetent patients can also be affected. This review proposes to evaluate cryptococcosis focusing on studies of this mycosis in Brazilian territory; moreover, recent advances in the understanding of its virulence mechanism, animal models in research are also assessed. For this, literature review as realized in PubMed, Scielo, and Brazilian legislation. In Brazil, cryptococcosis has been identified as one of the most lethal fungal infections among HIV patients and C. neoformans VNI and C. gattii VGII are the most prevalent genotypes. Moreover, different clinical settings published in Brazil were described. As in other countries, cryptococcosis is difficult to treat due to a limited therapeutic arsenal, which is highly toxic and costly. The presence of a polysaccharide capsule, thermo-tolerance, production of melanin, biofilm formation, mechanisms for iron use, and morphological alterations is an important virulence mechanism of these yeasts. The introduction of cryptococcosis as a compulsory notification disease could improve data regarding incidence and help in the management of these infections.

The Anti-Biofilm Efficacy of Caffeic Acid Phenethyl Ester (CAPE) In Vitro and a Murine Model of Oral Candidiasis.

Candida albicans is the main fungal species associated with the development of oral candidiasis. Currently, therapeutic options for these infections are limited by the adverse effects of antifungal drugs and by the emergence of drug resistant strains. Thus, the development of new antifungal agents is needed for the prevention and treatment of oral Candida infections. Caffeic acid phenethyl ester (CAPE) is a natural compound from propolis polyphenolic groups that exhibits many pharmacological properties. In this study, we investigated whether CAPE can have antifungal and immunomodulatory effects on oral candidiasis. Preliminary tests to assess the antifungal activity of CAPE were performed using the Minimum Inhibitory Concentration (MIC) assay that demonstrated inhibition in a range from 16 to 32 µg/mL, confirming its antifungal activity on several C. albicans strains isolated from the oral cavity. Subsequently, we analyzed Candida spp biofilms formed in vitro, in which CAPE treatment at 5 x MIC caused a reduction of 68.5% in the total biomass and ~2.60 Log in the viable cell count (CFU/mL) in relation to the untreated biofilm (p<0.0001). Next, RNA was extracted from untreated and CAPE-treated biofilms and analyzed by real-time qPCR. A series of genes analyzed (ALS1, ECE1, EPA1, HWP1, YWP1, BCR1, BGR1, CPH1, EFG1, NDT80, ROB1, TEC1, UME6, SAP2, SAP5, PBL2, and LIP9) were downregulated by CAPE compared to the untreated control group (p<0.0001). In in vivo studies using Galleria mellonella, the treatment with CAPE prolonged survival of larvae infected by C. albicans by 44.5% (p < 0.05) and accompanied by a 2.07-fold increase in the number of hemocytes. Flow cytometry revealed the most prominent increases were in types P2 and P3 hemocytes, granular cells, which phagocytize pathogens. In addition, CAPE treatment decreased the fungal load in the hemolymph and stimulated the expression of antifungal peptide genes such as galiomicin and gallerimycin. The antifungal and immunomodulatory activities observed in G. mellonella were extended to a murine model of oral candidiasis, in which CAPE decreased the levels of C. albicans colonization (~2 log CFU/mL) in relation to the untreated control group. In addition, CAPE treatment significantly reduced pseudomembranous lesions, invasion of hyphae on epithelium surfaces, tissue damage and inflammatory infiltrate (p < 0.05). CAPE was also able to increase the expression of ß-defensin 3 compared to the infected and untreated group by 3.91-fold (p < 0.0001). Taken together, these results show that CAPE has both antifungal and immunomodulatory effects, making it a promising natural antifungal agent for the treatment and prevention of candidiasis and shows impact to oral candidiasis.

Probiotic Effects of Lactobacillus paracasei 28.4 to Inhibit Streptococcus mutans in a Gellan-Based Formulation.

Streptococcus mutans is considered to be a major bacterium involved in dental caries, and the control of virulence mechanisms is fundamental to prevent disease. Probiotics present a promising preventive method; however, the use of probiotics requires its incorporation into delivery materials to facilitate oral colonization. Thus, we performed a comprehensive study examining preventive effects of Lactobacillus paracasei 28.4-enriched gellan hydrogel materials to inhibit S. mutans in planktonic and biofilm states, addressing its influence in the production of extracellular polysaccharides (EPS) and altered gene expression of several cariogenic virulence factors. L. paracasei 28.4, a strain isolated from the oral cavity of a caries-free individual, was incorporated in three gellan hydrogels (0.5%, 0.75%, and 1% w/v). The pretreatment with probiotic-gellan formulations provided a release of L. paracasei cells over 24 h that was sufficient to inhibit the planktonic growth of S. mutans, independent of the gellan concentrations and pH variations. This pretreatment also had inhibitory activity against S. mutans biofilms, exhibiting a reduction of 0.57 to 1.54 log10 in CFU/mL (p < 0.0001) and a decrease of 68.8 to 71.3% in total biomass (p < 0.0001) compared with the control group. These inhibitory effects were associated with the decreased production of EPS by 80% (p < 0.0001) and the downregulation of luxS, brpA, gbpB, and gtfB genes. The gellan formulation containing L. paracasei 28.4 exhibited probiotic effects for preventing S. mutans growth, biofilm formation, and production of cariogenic factors to suggest possible use in tooth decay prevention.

The Postbiotic Activity of Lactobacillus paracasei 28.4 Against Candida auris.

Candida auris has emerged as a medically important pathogen with considerable resistance to antifungal agents. The ability to produce biofilms is an important pathogenicity feature of this species that aids escape of host immune responses and antimicrobial agents. The objective of this study was to verify antifungal action using in vitro and in vivo models of the Lactobacillus paracasei 28.4 probiotic cells and postbiotic activity of crude extract (LPCE) and fraction 1 (LPF1), derived from L. paracasei 28.4 supernatant. Both live cells and cells free supernatant of L. paracasei 28.4 inhibited C. auris suggesting probiotic and postbiotic effects. The minimum inhibitory concentration (MIC) for LPCE was 15 mg/mL and ranges from 3.75 to 7.5 mg/mL for LPF1. Killing kinetics determined that after 24 h treatment with LPCE or LPF1 there was a complete reduction of viable C. auris cells compared to fluconazole, which decreased the initial inoculum by 1-logCFU during the same time period. LPCE and LPF1 significantly reduced the biomass (p = 0.0001) and the metabolic activity (p = 0.0001) of C. auris biofilm. There was also a total reduction (~108 CFU/mL) in viability of persister C. auris cells after treatment with postbiotic elements (p < 0.0001). In an in vivo study, injection of LPCE and LPF1 into G. mellonella larvae infected with C. auris prolonged survival of these insects compared to a control group (p < 0.05) and elicited immune responses by increasing the number of circulating hemocytes and gene expression of antimicrobial peptide galiomicin. We concluded that the L. paracasei 28.4 cells and postbiotic elements (LPCE and LPF1) have antifungal activity against planktonic cells, biofilms, and persister cells of C. auris. Postbiotic supplementation derived from L. paracasei 28.4 protected G. mellonella infected with C. auris and enhanced its immune status indicating a dual function in modulating a host immune response.

Candida Biofilms: An Update on Developmental Mechanisms and Therapeutic Challenges.

Fungi of the genus Candida are important etiological agents of superficial and life-threatening infections in individuals with a compromised immune system. One of the main characteristics of Candida is its ability to form highly drug tolerance biofilms in the human host. Biofilms are a dynamic community of multiple cell types whose formation over time is orchestrated by a network of transcription regulators. In this brief review, we provide an update of the processes involved in biofilm formation by Candida spp. (formation, treatment, and control), as well as the transcriptional circuitry that regulates its development and interactions with other microorganisms. Candida albicans is known to build mixed species biofilms with other Candida species and with various other bacterial species in different host niches. Taken together, these properties play a key role in Candida pathogenesis. In addition, this review gathers recent studies with new insights and perspectives for the treatment and control of Candida biofilms.

Development of Probiotic Formulations for Oral Candidiasis Prevention: Gellan Gum as a Carrier To Deliver Lactobacillus paracasei 28.4.

Probiotics might provide an alternative approach for the control of oral candidiasis. However, studies on the antifungal activity of probiotics in the oral cavity are based on the consumption of yogurt or other dietary products, and it is necessary to use appropriate biomaterials and specific strains to obtain probiotic formulations targeted for local oral administration. In this study, we impregnated gellan gum, a natural biopolymer used as a food additive, with a probiotic and investigated its antifungal activity against Candida albicansLactobacillus paracasei 28.4, a strain recently isolated from the oral cavity of a caries-free individual, was incorporated in several concentrations of gellan gum (0.6% to 1% [wt/vol]). All tested concentrations could incorporate L. paracasei cells while maintaining bacterial viability. Probiotic-gellan gum formulations were stable for 7 days when stored at room temperature or 4°C. Long-term storage of bacterium-impregnated gellan gum was achieved when L. paracasei 28.4 was lyophilized. The probiotic-gellan gum formulations provided a release of L. paracasei cells over 24 h that was sufficient to inhibit the growth of C. albicans, with effects dependent on the cell concentrations incorporated into gellan gum. The probiotic-gellan gum formulations also had inhibitory activity against Candida sp. biofilms by reducing the number of Candida sp. cells (P < 0.0001), decreasing the total biomass (P = 0.0003), and impairing hyphae formation (P = 0.0002), compared to the control group which received no treatment. Interestingly, a probiotic formulation of 1% (wt/vol) gellan gum provided an oral colonization of L. paracasei in mice with approximately 6 log CFU/ml after 10 days. This formulation inhibited C. albicans growth (P < 0.0001), prevented the development of candidiasis lesions (P = 0.0013), and suppressed inflammation (P = 0.0006) compared to the mice not treated in the microscopic analysis of the tongue dorsum. These results indicate that gellan gum is a promising biomaterial and can be used as a carrier system to promote oral colonization for probiotics that prevent oral candidiasis.

Effects of surface pre-reacted glass-ionomer (S-PRG) eluate on Candida spp.: antifungal activity, anti-biofilm properties, and protective effects on Galleria mellonella against C. albicans infection.

Surface pre-reacted glass-ionomer (S-PRG) is a bioactive filler produced by PRG technology, which is applied to various dental materials. The inhibitory effects of S-PRG eluate against Candida, the most common fungal oral pathogen, were investigated. Minimum inhibitory concentrations (MIC) and anti-biofilm activities were tested against Candida albicans, Candida glabrata, Candida krusei, and Candida tropicalis. For the in vivo study, Galleria mellonella was used as a model to evaluate the effects of S-PRG on toxicity, hemocyte counts and candidiasis. The MIC of S-PRG ranged from 5 to 40% (v/v). S-PRG eluate exhibited anti-biofilm activity for all the Candida species tested. Furthermore, injection of S-PRG eluate into G. mellonella was not toxic to the larvae and protected G. mellonella against experimental candidiasis. In addition, S-PRG eluate inhibited biofilm formation by C. albicans, C. glabrata, C. krusei, and C. tropicalis and exerted protective effects on G. mellonella against experimental candidiasis in vivo.

Two sporulated Bacillus enhance immunity in Galleria mellonella protecting against Candida albicans.

The aim of this study was to evaluate the effects of Bacillus subtilis and Bacillus atrophaeus on Galleria mellonella immunity challenged by Candida albicans. Firstly, we analyzed the susceptibility of G. mellonella to bacilli (vegetative and sporulating forms). It was found that both vegetative and sporulating forms were not pathogenic to G. mellonella at a concentration of 1 × 104 cells/larva. Next, larvae were pretreated with two species of Bacillus, in the vegetative and sporulating forms, and then challenged with C. albicans. In addition, the gene expression of antimicrobial peptides (AMPs) such as Gallerimycin, Gloverin, Cecropin-D and Galiomicin was investigated. Survival rates increased in the Bacillus treated larvae compared with control larvae inoculated with C. albicans only. Cells and spores of Bacillus spp. upregulated Gloverin, Galiomicin and Gallerimycin genes in relation to the control group (PBS + PBS). When these larvae were infected with C. albicans, the group pretreated with spores of B. atrophaeus and B. subtilis showed a greater increase in expression of Galiomycin (49.08-fold and 13.50-fold) and Gallerimycin (27.88-fold and 68.15-fold), respectively, compared to the group infected with C. albicans only (p = 0.0001). After that, we investigated the effects of B. subtilis and B. atrophaeus on immune system of G. mellonella evaluating the number of hemocytes, quantification of melanization, cocoon formation and colony forming units (CFU) count. Hemocyte count increased in response to stimulation by Bacillus, and a higher increase was achieved when larvae were inoculated with B. subtilis spores (p = 0.0011). In the melanization assay, all groups tested demonstrated lower production of melanin compared to that in the phosphate-buffered saline (PBS) group. In addition, full cocoon formation was observed in all groups analyzed, which corresponded to a healthier wax worm. Hemolymph culture revealed higher growth of B. atrophaeus and B. subtilis in the groups inoculated with spores. We concluded that spores and cells of B. atrophaeus and B. subtilis stimulated the immune system of G. mellonella larvae and protected them of C. albicans infection.

Galleria mellonella as an experimental model to study human oral pathogens.

The oral cavity is home to a microbial community of more than 800 species. This important microbiome is formed by commensal and opportunistic bacteria, fungi and viruses. Several distinct habitats within the mouth support heterogeneous microbial communities that constitute an important link between oral and general health. The use of animal models for in vivo studies in microbial pathogenicity is well established in the scientific community. Galleria mellonella as a model host has increased in use significantly in the last few years. This invertebrate model provides studies on a large scale, serving as screens for studies on vertebrate animals, such as mice and rats. In this review, different studies of microbial genera of dental importance (Enterococcus, Candida, Lactobacillus, Porphyromonas and Streptococcus) are discussed, highlighting the use of G. mellonella as a suitable model for studying pathogenesis, efficacy of antimicrobial compounds, and immune responses.

Lactobacillus species increase the survival of Galleria mellonella infected with Candida albicans and non-albicans Candida clinical isolates.

Investigation into new therapeutic strategies, such as the use of bacterial isolates with probiotic characteristics, has increased in importance due to the high incidence of Candida albicans and non-albicans Candida infections. This study evaluates Lactobacillus paracasei, Lactobacillus fermentum and Lactobacillus rhamnosus strains as prophylactic and therapeutic agents against infection caused by Candida albicans, Candida glabrata, Candida krusei, and Candida tropicalis in a Galleria mellonella model. Prophylactic treatment provided greater benefits during Candida spp. infection, increasing G. mellonella survival, compared to therapeutic treatment. This study demonstrated that the different Lactobacillus species are potent prophylactic agents of Candida species infection.

Effects of Bacillus subtilis on Candida albicans: biofilm formation, filamentation and gene expression / Efeitos de Bacillus subtilis sobre Candida albicans: formação de biofilme, filamentação e expressão gênica

Objective: The aim of this study was evaluate the effect of Bacillus subtilis on Candida albicans biofilm formation and filamentation by evaluating the gene expression of ALS3, HWP1, BCR1, EFG1 and TEC1. Material and Methods: Mixed (C. albicans / B.subtilis) and monotypic biofilms were cultured in plates at 37°C for 48 h under shaking for counting viable cells (CFU / mL) and analysis of gene expression by real-time PCR. The C. albicans filamentation assay was performed in medium containing 10% fetal bovine serum at 37°C for 6 hours. Data was analysed by t-Student and Mann­ Whitney tests. Results: B. subtilis reduced the biofilm formation of C. albicans in 1 log when cultured in the same environment (p<0.0001). In addition, it significantly reduced the yeast - hypha transition affecting the morphology of C. albicans. Among all of the analyzed genes, the ALS3 and HWP1 genes were the most affected, achieving 111.1- and 333.3- fold decreases in the C. albicans biofilms associated with B. subtilis, respectively. Conclusion: B. subtilis reduced the biofilm formation and filamentation of C. albicans by negatively regulating the ALS3, HWP1, BCR1, EFG1 and TEC1 genes that are essential for the production of biofilm and hyphae. (AU)

Objetivo: O objetivo deste estudo foi avaliar o efeito de Bacillus subtilis sobre a formação de biofilme e filamentação de Candida albicans através da avaliação da expressão dos genes ALS3, HWP1, BCR1, EFG1 and TEC1. Material e métodos: Biofilmes monotípicos e mistos (C. albicans / B.subtilis) foram cultivados em placas a 37°C por 48 h sob agitação, para a contagem de células viáveis (UFC/mL) e para a análise da expressão gênica por PCR em tempo real. O ensaio de filamentação de C. albicans foi realizado em meio contendo 10% de soro fetal bovino a 37°C por 6 h. Os dados obtidos foram analisados por testes t-Student e Mann­Whitney. Resultados: B.subtilis reduziu em 1 log a formação de biofilme por C. albicans quando cultivados no mesmo ambiente (p<0.0001). Além disso, reduziu significantemente a transição de levedura para hifa, afetando assim, a morfologia de C. albicans. Em relação aos genes analisados, os genes ALS3 e HWP1 foram os mais regulados negativamente, com uma diminuição de 111,1 e 333,3 vezes, respectivamente, na sua expressão em biofilmes de C. albicans associados a B. subtilis. Conclusão: B. subtilis reduziu a filamentação e a formação de biofilme de C. albicans através da regulação negativa dos genes ALS3, HWP1, BCR1, EFG1 e TEC1, que são essenciais na produção de hifas e de biofilme. (AU)

Recent Advances in the Use of Galleria mellonella Model to Study Immune Responses against Human Pathogens.

The use of invertebrates for in vivo studies in microbiology is well established in the scientific community. Larvae of Galleria mellonella are a widely used model for studying pathogenesis, the efficacy of new antimicrobial compounds, and immune responses. The immune system of G. mellonella larvae is structurally and functionally similar to the innate immune response of mammals, which makes this model suitable for such studies. In this review, cellular responses (hemocytes activity: phagocytosis, nodulation, and encapsulation) and humoral responses (reactions or soluble molecules released in the hemolymph as antimicrobial peptides, melanization, clotting, free radical production, and primary immunization) are discussed, highlighting the use of G. mellonella as a model of immune response to different human pathogenic microorganisms.

Galleria mellonella experimental model for bat fungal pathogen Pseudogymnoascus destructans and human fungal pathogen Pseudogymnoascus pannorum.

Laboratory investigations of the pathogenesis of Pseudogymnoascus destructans, the fungal causal agent of bat White Nose Syndrome (WNS), presents unique challenges due to its growth requirements (4°-15°C) and a lack of infectivity in the current disease models. Pseudogymnoascus pannorum is the nearest fungal relative of P. destructans with wider psychrophilic - physiological growth range, and ability to cause rare skin infections in humans. Our broad objectives are to create the molecular toolkit for comparative study of P. destructans and P. pannorum pathogenesis. Towards these goals, we report the successful development of an invertebrate model in the greater wax moth Galleria mellonella. Both P. destructans and P. pannorum caused fatal disease in G. mellonella and elicited immune responses and histopathological changes consistent with the experimental disease.

Inhibitory effect of probiotic Lactobacillus supernatants from the oral cavity on Streptococcus mutans biofilms.

Probiotics can release bioactive substances that can inhibit the growth and biofilm formation of pathogenic microorganisms such as Streptococcus mutans. In this context, we evaluated whether the supernatants of Lactobacillus strains isolated from caries-free subjects can inhibit S. mutans, one of the most important bacteria for dental caries. First, the supernatants of 22 Lactobacillus strains were screened for antibacterial activity against S. mutans in planktonic cultures. All 22 Lactobacillus strains studied (100%) showed antibacterial activity. Thereafter, the Lactobacillus strains with the greatest reductions in the planktonic S. mutans cultures were tested on biofilms. The L. fermentum 20.4, L. paracasei 11.6, L. paracasei 20.3 and L. paracasei 25.4 strains could significantly reduce the number of S. mutans cells in biofilms formed in hydroxyapatite (p < 0.05). This reduction was also confirmed by scanning electron microscopy analysis and was not caused by the decreased pH value in the medium (p > 0.05). In addition, the supernatants of these probiotic strains could also reduce the total biomass of S. mutans biofilms (p < 0.05). In conclusion, most of the Lactobacillus strains tested have some antibacterial activity against S. mutans. L. fermentum 20.4, L. paracasei 11.6, L. paracasei 20.3 and L. paracasei 25.4 produce bioactive substances that caused a significant reduction in S. mutans biofilms.

Candida tropicalis affects the virulence profile of Candida albicans: an in vitro and in vivo study.

Candida albicans and Candida tropicalis are commensal microorganisms occurring in the oral cavity of approximately 50%-70% of healthy individuals. However, these microbes can become pathogenic through changes in the environment or weakened host immune system. Thus, the aim of this investigation was to evaluate the interaction between species of the genus Candida in the biofilm formation, filamentation, gene expression and virulence in Galleria mellonella. Coincubation of C. albicans with C. tropicalis cells after 48 h resulted in significant reduction of biofilm formation by decreasing viable cell counts, metabolic activity and hyphal growth. The C. albicans genes (BCR1, CPH1, EFG1, UME6, HWP1, ALS3, SAP5 and PLB2) were quantified by quantitative real-time polymerase chain reaction and most of genes were downregulated. Regarding in vivo assay, the groups that the larvae received C. albicans and C. tropicalis had a significant survival increase compared to the control group of C. albicans (P = 0.0001) in agreement with the in vitro results. In conclusion, C. tropicalis colonization was associated with a decrease in the growth of C. albicans, suggesting an antagonistic relation between these two species. Therefore, C. tropicalis by reducing C. albicans virulence profile may limit the ability of this pathogenic fungus to cause infection.

Lactobacillus paracasei 28.4 reduces in vitro hyphae formation of Candida albicans and prevents the filamentation in an experimental model of Caenorhabditis elegans.

The objective of this study was to evaluate the influence of microbe-microbe interactions to identify a strain of Lactobacillus that could reduce the filamentation of Candida albicans ATCC 18804 using in vitro and in vivo models. Thus presenting a probiotic effect against the fungal pathogen. First, we analyzed the ability of 25 clinical isolates of Lactobacillus to reduce filamentation in C. albicans in vitro. We found that L. paracasei isolate 28.4 exhibited the greatest reduction of C. albicans hyphae (p = 0.0109). This reduction was confirmed by scanning electron microscopy analysis. The influence of C. albicans filamentation was found to be contributed through reduced gene expression of filament associated genes (TEC1 and UME6). In an in vivo study, prophylactic provisions with L. paracasei increased the survival of Caenorhabditis elegans worms infected with C. albicans (p = 0.0001) by 29%. Prolonged survival was accompanied by the prevention of cuticle rupture of 27% of the worms by filamentation of C. albicans, a phenotype that is characteristic of C. albicans killing of nematodes, compared to the control group. Lactobacillus paracasei isolate 28.4 reduced the filamentation of C. albicans in vitro by negatively regulating the TEC1 and UME6 genes that are essential for the production of hyphae. Prophylactic provision of Lactobacillus paracasei 28.4 protected C. elegans against candidiasis in vivo. L. paracasei 28.4 has the potential to be employed as an alternative method to control candidiasis.

Antifungal activity of clinical Lactobacillus strains against Candida albicans biofilms: identification of potential probiotic candidates to prevent oral candidiasis.

This study isolated Lactobacillus strains from caries-free subjects and evaluated the inhibitory effects directly on three strains of C. albicans, two clinical strains and one reference strain. Thirty Lactobacillus strains were isolated and evaluated for antimicrobial activity against in vitro C. albicans biofilms. L. paracasei 28.4, L. rhamnosus 5.2 and L. fermentum 20.4 isolates exhibited the most significant inhibitory activity against C. albicans. Co-incubation between these microorganisms resulted in deterrence of biofilm development and retardation of hyphal formation. The hindrance of biofilm development was characterized by the downregulated expression of C. albicans biofilm-specific genes (ALS3, HWP1, EFG1 and CPH1). L. paracasei 28.4, L. rhamnosus 5.2 and L. fermentum 20.4 demonstrated the ability to exert antifungal activity through the inhibition of C. albicans biofilms.

Evaluation of the maladaptation of the test base in acrylic resin regarding the technique of preparation, place of measurement and storage time / Avaliação da desadaptação da base de prova em resina acrílica em relação à técnica de confecção, local de mensuração e tempo de armazenagem

Introduction: The most used material for the preparation of the baseplates is the acrylic resin, but it can present distortions. Objective: To evaluate preparation technique, region and storage time that presents less maladaptation of the base when made with self-cured acrylic resin. Material and method: Models were created in gypsum type III simulating edentulous maxilla, as divided into 3 groups (n = 10): GC (control group) thermopolymerizable acrylic resin; G1 - manual adaptation technique and G2 - drip technique. For the measurements, silicone by condensation of light consistency that was interposed between base and model was used. With a hydraulic press, 50 kg pressure was applied leading the base of the model. The obtained mold was measured in the palate, canine and molar regions with a digital caliper at the following times: immediately after the base polymerization, at 24, 48, 72, 96 hours and one week. The results were submitted to statistical analysis. Result: G1 presented maladaptation of 0.43 mm ± 0.10, while G2 obtained 0.39 mm ± 0.11. The lowest maladaptation occurred in the CG. The palate region presented greater maladaptation (0.52 ± 0.07) and the canine region, the lowest (CD = 0.27 mm ± 0.07 and CE = 0.27 ± 0.09); There was no statistically significant difference for storage times. Conclusion: G2 presented lower values than G1, with no statistically significant difference. The palate region presented greater maladaptation, followed by molars and canines. The bases continued to maladaptation the model after the immediate polymerization, with no statistically significant difference.

Introdução: O material mais empregado para confecção da base de prova é a resina acrílica por oferecer maior rapidez e praticidade, embora tenda a maior distorção. Objetivo: Avaliar técnica de confecção, região e tempo de armazenagem que apresente menor desadaptação da base de prova confeccionada com resina acrílica ativada quimicamente. Material e método: Confeccionaram-se modelos em gesso tipo III simulando maxila edêntula que foram divididos em 3 grupos (n = 10): GC - (grupo controle) resina acrílica termopolimerizável; G1 - técnica da adaptação manual; e G2 - técnica do gotejamento. Para as mensurações utilizou-se silicone por condensação de consistência leve que foi interposto entre base e modelo. Com uma prensa hidráulica aplicou-se pressão de 50 kg levando a base de encontro ao modelo. O molde obtido foi mensurado nas regiões de palato, caninos e molares com paquímetro digital nos seguintes tempos: imediatamente após a polimerização da base, em 24, 48, 72, 96 horas e uma semana. Os resultados foram submetidos à análise estatística. Resultado: O G1 apresentou média de desadaptação de 0,43mm±0,10, enquanto o G2 obteve 0,39 mm ± 0,11. Os menores valores de desadaptação ocorreram no GC; A região do palato apresentou maior desadaptação (0,52 mm ± 0,07) e a região de caninos, as menores (CD = 0,27 mm ± 0,07 e CE = 0,27 ± 0,09); Não houve diferença estatisticamente significante para os tempos de armazenagem. Conclusão: O G2 apresentou menores valores que o G1, sem diferença estatisticamente significante; A região de palato apresentou maior desadaptação, seguida de molares e caninos; As bases continuaram desadaptando ao modelo após a polimerização imediata, sem diferença estatisticamente significante.