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1.
Front Cell Infect Microbiol ; 12: 895068, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35646731

RESUMO

Candida albicans Als1 is a large cell-surface glycoprotein most often discussed for its role in mediating ligand-binding and aggregative interactions. Relative to a wild-type control, deletion of ALS1 produced a strain that showed delayed germ-tube formation and delayed disease progression in a murine model of disseminated candidiasis. Populations of Δals1/Δals1 cultured cells had a higher proportion of smaller cells compared to wild-type or ALS1 reintegrant control cultures. The goal of this work was to investigate whether this difference in cell-size distributions was responsible for delayed germ-tube formation and delayed disease progression. Flow cytometry was used to select populations of wild-type and Δals1/Δals1 cells with varied cell-size distributions. Delayed germ-tube formation was demonstrated for small cells sorted from a wild-type (ALS1/ALS1) culture population. Large cells sorted from a Δals1/Δals1 culture formed germ tubes as quickly as the wild-type control demonstrating clearly that the Δals1/Δals1 germ-tube formation delays were attributable to cell size. In vivo, smaller-sized cells of the wild-type control showed fewer colony-forming units (cfu) per gram of kidney tissue and less-severe histopathology lesions compared to larger cells of the same strain. The Δals1/Δals1 strain showed reduced cfu/g of kidney tissue and less-severe lesions compared to the wild-type control. However, isolation and testing of the larger cells from the Δals1/Δals1 population increased cfu/g of tissue and showed increased lesion severity compared to the overall mutant cell population. In vivo hypha lengths from the large, sorted Δals1/Δals1 cells were comparable to those for the wild-type control strain. These results demonstrated that a large share of the Δals1/Δals1 in-vivo phenotype was attributable to cell size. Collectively, the data suggest a role for Als1 in C. albicans cell size homeostasis, a novel hypothesis for further exploration.


Assuntos
Candida albicans , Candidíase , Esclerose Amiotrófica Lateral , Animais , Candida albicans/genética , Progressão da Doença , Proteínas Fúngicas/genética , Hifas , Camundongos
2.
PLoS Pathog ; 18(6): e1010089, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35687592

RESUMO

Whether to commit limited cellular resources toward growth and proliferation, or toward survival and stress responses, is an essential determination made by Target of Rapamycin Complex 1 (TORC1) for a eukaryotic cell in response to favorable or adverse conditions. Loss of TORC1 function is lethal. The TORC1 inhibitor rapamycin that targets the highly conserved Tor kinase domain kills fungal pathogens like Candida albicans, but is also severely toxic to human cells. The least conserved region of fungal and human Tor kinases are the N-terminal HEAT domains. We examined the role of the 8 most N-terminal HEAT repeats of C. albicans Tor1. We compared nutritional- and stress responses of cells that express a message for N-terminally truncated Tor1 from repressible tetO, with cells expressing wild type TOR1 from tetO or from the native promoter. Some but not all stress responses were significantly impaired by loss of Tor1 N-terminal HEAT repeats, including those to oxidative-, cell wall-, and heat stress; in contrast, plasma membrane stress and antifungal agents that disrupt plasma membrane function were tolerated by cells lacking this Tor1 region. Translation was inappropriately upregulated during oxidative stress in cells lacking N-terminal Tor1 HEAT repeats despite simultaneously elevated Gcn2 activity, while activation of the oxidative stress response MAP kinase Hog1 was weak. Conversely, these cells were unable to take advantage of favorable nutritional conditions by accelerating their growth. Consuming oxygen more slowly than cells containing wild type TOR1 alleles during growth in glucose, cells lacking N-terminal Tor1 HEAT repeats additionally were incapable of utilizing non-fermentable carbon sources. They were also hypersensitive to inhibitors of specific complexes within the respiratory electron transport chain, suggesting that inefficient ATP generation and a resulting dearth of nucleotide sugar building blocks for cell wall polysaccharides causes cell wall integrity defects in these mutants. Genome-wide expression analysis of cells lacking N-terminal HEAT repeats showed dysregulation of carbon metabolism, cell wall biosynthetic enzymes, translational machinery biosynthesis, oxidative stress responses, and hyphal- as well as white-opaque cell type-associated genes. Targeting fungal-specific Tor1 N-terminal HEAT repeats with small molecules might selectively abrogate fungal viability, especially when during infection multiple stresses are imposed by the host immune system.


Assuntos
Candida albicans , Proteínas Fúngicas , Candida albicans/metabolismo , Carbono/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Humanos , Hifas , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismo
3.
Sci Total Environ ; 837: 155498, 2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-35523342

RESUMO

The effects of mycorrhiza and its external hyphae on the response of soil microbes to global warming remain unclear. This study investigates the role of mycorrhiza and its hyphae in regulating soil microbial community under warming by examining the microbial biomass and composition in the ingrowth cores of arbuscular mycorrhiza (AM) plant, Fargesia nitida, and ectomycorrhiza (ECM) plant, Picea asperata, with/without mycorrhiza/hyphae and experimental warming. The results showed that warming significantly increased the biomass of all soil microbes (by 19.89%-137.48%) and altered the microbial composition in both plant plots without mycorrhiza/hyphae. However, this effect was weakened in the presence of mycorrhiza or hyphae. In F. nitida plots, warming did not significantly affect biomass and composition of most soil microbial groups when mycorrhiza or hyphae were present. In P. asperata plots, warming significantly increased the total and ECM fungi (ECMF) biomass in the presence of hyphae (p < 0.05) and the total, Gn, and AM fungi (AMF) biomass in the presence of mycorrhiza (p < 0.05). Meanwhile, the response of enzyme activities to warming was also altered with mycorrhiza or hyphae. Additionally, soil microbial community composition was mainly influenced by soil available phosphorus (avaP), while enzyme activities depended on soil avaP, dissolved organic carbon (DOC), and nitrate concentrations. Our results indicate that mycorrhiza and its hyphae are essential in regulating the response of microbes to warming.


Assuntos
Microbiota , Micorrizas , Biomassa , Carbono , Hifas , Micorrizas/fisiologia , Fósforo , Plantas , Solo , Microbiologia do Solo , Tibet
4.
Acta Biomater ; 145: 272-282, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35421618

RESUMO

Little is known about the mechanical and material properties of hyphae, the single constituent material of Agaricomycetes fungi, despite a growing interest in fungus-based materials. In the Agaricomycetes (the mushrooms and allies), there are three types of hyphae that make up sporocarps: generative, skeletal, and ligative. All filamentous Agaricomycetes can be categorized into one of three categories of hyphal systems that compose them: monomitic, dimitic, and trimitic. Monomitic systems have only generative hyphae. Dimitic systems have generative and either skeletal (most common) or ligative. Trimitic systems are composed of all three kinds of hyphae. SEM imaging, compression testing, and theoretical modeling were used to characterize the material and mechanical properties of representative monomitic, dimitic, and trimitic sporocarps. Compression testing revealed an increase in the compression modulus and compressive strength with the addition of more hyphal types (monomitic to dimitic and dimitic to trimitic). The mesostructure of the trimitic sporocarp was tested and modeled, suggesting that the difference in properties between the solid material and the microtubule mesostructure is a result of differences in structure and not material. Theoretical modeling was completed to estimate the mechanical properties of the individual types of hyphae and showed that skeletal hyphae make the largest contribution to mechanical properties of fungal sporocarps. Understanding the contributions of the different types of hyphae may help in the design and application of fungi-based or bioinspired materials. STATEMENT OF SIGNIFICANCE: This research studies the material and mechanical properties of fungal sporocarps and their hyphae, the single constituent material of Agaricomycetes fungi. Though some work has been done on fungal hyphae, this research studies hyphae in context of the three hyphal systems found in Agaricomycetes fungi and estimates the properties of the hyphal filaments, which has not been done previously. This characterization was performed by analyzing the structures and mechanical properties of fungal sporocarps and calculating the theoretical mechanical properties of their hyphae. This data and the resulting conclusions may lead to a better design and implementation process of fungi-based materials in various applications using the properties now known or calculated.


Assuntos
Citoesqueleto , Hifas , Microtúbulos
5.
Sci Rep ; 12(1): 5874, 2022 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-35393437

RESUMO

Species in the Fusarium solani species complex are fast growing, environmental saprophytic fungi. Members of this genus are filamentous fungi with a wide geographical distribution. Fusarium keratoplasticum and F. falciforme have previously been isolated from sea turtle nests and have been associated with high egg mortality rates. Skin lesions were observed in a number of stranded, post-hatchling loggerhead sea turtles (Caretta caretta) in a rehabilitation facility in South Africa. Fungal hyphae were observed in epidermal scrapes of affected turtles and were isolated. The aim of this study was to characterise the Fusarium species that were isolated from these post-hatchling loggerhead sea turtles (Caretta caretta) that washed up on beaches along the South African coastline. Three gene regions were amplified and sequenced, namely the internal transcribed spacer region (ITS), a part of the nuclear large subunit (LSU), and part of the translation elongation factor 1 α (tef1) gene region. Molecular characteristics of strains isolated during this study showed high similarity with Fusarium isolates, which have previously been associated with high egg mortality rates in loggerhead sea turtles. This is the first record of F. keratoplasticum, F. falciforme and F. crassum isolated from stranded post-hatchling loggerhead sea turtles in South Africa.


Assuntos
Fusarium , Tartarugas , Animais , Fusarium/genética , Hifas , África do Sul , Tartarugas/microbiologia
6.
Microbes Environ ; 37(6)2022.
Artigo em Inglês | MEDLINE | ID: mdl-35387945

RESUMO

Filamentous fungi grow by the elongation of tubular cells called hyphae and form mycelia through repeated hyphal tip growth and branching. Since hyphal growth is closely related to the ability to secrete large amounts of enzymes or invade host cells, a more detailed understanding and the control of its growth are important in fungal biotechnology, ecology, and pathogenesis. Previous studies using fluorescence imaging revealed many of the molecular mechanisms involved in hyphal growth. Raman microspectroscopy and imaging methods are now attracting increasing attention as powerful alternatives due to their high chemical specificity and label-free, non-destructive properties. Spatially resolved information on the relative abundance, structure, and chemical state of multiple intracellular components may be simultaneously obtained. Although Raman studies on filamentous fungi are still limited, this review introduces recent findings from Raman studies on filamentous fungi and discusses their potential use in the future.


Assuntos
Fungos , Hifas , Citoplasma , Análise Espectral
7.
Mycorrhiza ; 32(3-4): 269-280, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35419710

RESUMO

Achlorophyllous, mycoheterotrophic plants often have an elaborate mycorrhizal colonization pattern, allowing a sustained benefit from external fungal root penetrations. The present study reveals the root anatomy and mycorrhizal pattern of eight mycoheterotrophic Thismia spp. (Thismiaceae), all of which show separate tissue compartments segregating different hyphal shapes of the mycorrhizal colonization, as there are intact straight, coiled and peculiarly knotted hyphae as well as degenerated clumps of hyphal material. Those tissue compartments in Thismia roots potentially comprise exo-, meso- and endoepidermae, and exo-, meso- and endocortices, although not all species develop all these root layers. Differences in details among species according to anatomy (number of root layers, cell sizes and shapes) and colonization pattern (hyphal shapes within cells) are striking and can be discussed as an evolutionary series towards increasing mycorrhizal complexity which roughly parallels the recently established phylogeny of Thismia. We suggest functional explanations for why the distinct elements of the associations can contribute to the mycorrhizal advantage for the plants and, thus, we emphasize the relevance of structural traits for mycorrhizae.


Assuntos
Micorrizas , Evolução Biológica , Hifas , Raízes de Plantas/microbiologia , Sementes , Simbiose
9.
Front Cell Infect Microbiol ; 12: 855229, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35392604

RESUMO

Candida sp. are among the most common fungal commensals found in the human microbiome. Although Candida can be found residing harmlessly on the surface of the skin and mucosal membranes, these opportunistic fungi have the potential to cause superficial skin, nail, and mucus membrane infections as well as life threatening systemic infections. Severity of infection is dependent on both fungal and host factors including the immune status of the host. Virulence factors associated with Candida sp. pathogenicity include adhesin proteins, degradative enzymes, phenotypic switching, and morphogenesis. A central transcriptional regulator of morphogenesis, the transcription factor Efg1 was first characterized in Candida albicans in 1997. Since then, EFG1 has been referenced in the Candida literature over three thousand times, with the number of citations growing daily. Arguably one of the most well studied genes in Candida albicans, EFG1 has been referenced in nearly all contexts of Candida biology from the development of novel therapeutics to white opaque switching, hyphae morphology to immunology. In the review that follows we will synthesize the research that has been performed on this extensively studied transcription factor and highlight several important unanswered questions.


Assuntos
Candida albicans , Proteínas de Ligação a DNA , Proteínas Fúngicas , Fatores de Transcrição , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Humanos , Hifas , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
10.
Mycologia ; 114(3): 533-543, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35482536

RESUMO

Most cytology reports on ectomycorrhiza (ECM) have been less focused on the fungal structures, especially on the exploratory organ. This study aimed to evaluate the morphological response of explorative mycelia of ECM forming fungi with and without the occurrence of the ECM host. We assessed the mycelial aggregates of Rhizopogon roseolus which was inoculated with and without Pinus thunbergii under controlled laboratory conditions. The mycelial aggregates with the host produced strikingly complex hyphal strands relative to those without host. Light microscopy revealed that the cytology and plectology of both mycelial aggregates had the approximately similar architecture. The tubular hyphae diameter without the host was consistently larger than that with the host. This study confirmed that the septa diameter of the tubular hyphae conjunction of mycelial aggregates with the host were shorter than those without the host. In addition, partially and completely dissolved septa of the tubular hyphae were evident with and without host. We also described the thromboplerous hyphae, which have rarely been reported in vitro. These hyphae were produced in higher numbers near the fungal inoculum with and without the host. The current findings will add to the current knowledge regarding ECM fungi mycelial aggregate response to the host.


Assuntos
Basidiomycota , Pinus , Hifas , Micélio , Pinus/microbiologia
11.
Chemosphere ; 300: 134526, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35398069

RESUMO

Manipulating the elemental composition is one major strategy to tune the properties of biochars to endow different functions to meet various application requirements. Compared with the widely reported plants- and manure-based precursors for biochars, microbes-based precursors take the lead in regional independency, rapid growth, morphology uniformity and abundant species with different adjustable elemental composition. In this work, fungal hyphae with massive microtubule structure were selected as a typical microbe precursor to prepare biochars whose catalysis capability was further evaluated by a representative reduction reaction of 4-nitrophenol (4-NP) using sodium borohydride as reductant. By simply increasing the nitrogen concentration in the culturing medium, the fungal hyphae derived biochars with increased nitrogen contents (2.1 wt% → 4.3 wt%) were successfully obtained after pyrolysis, showing almost two times higher catalysis ability (4.75 × 10-2 s-1 → 7.26 × 10-2 s-1) towards 4-nitrophenol. The Arrhenius equation calculation further proved that the more doping of nitrogen would increase the active sites rather than altering the reaction pathway. A high surface area of 997 m2 g-1 at pyrolysis temperature of 800 °C was obtained resulting from the fine microstructure of fungal hyphae. Higher pyrolysis temperature derived biochars remarkably promote their catalysis ability. These results indicated that biochars with controllable nitrogen contents can be prepared by cooperating culturing and pyrolysis processes, which pointed out an easy, rapid, scalable, and promising way to synthesis biochars with tunable functions for different applications.


Assuntos
Nitrogênio , Pirólise , Adsorção , Catálise , Carvão Vegetal/química , Hifas , Nitrogênio/química , Nitrofenóis , Temperatura
12.
Microbiol Spectr ; 10(2): e0188021, 2022 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-35234488

RESUMO

The role of endocytosis in Candida albicans secretion, filamentation, and virulence remains poorly understood, despite its importance as a fundamental component of intracellular trafficking. Given that secretory mutants display defects in endocytosis, we have focused our attention on endocytic mutants to understand the interconnection between endocytosis and other secretory pathways. Using a reverse-genetic approach based upon CRISPR-Cas9 mediated gene deletion, we studied the functions of the gene END3, which plays a key role in clathrin-based endocytosis. In the end3Δ/Δ null mutant, clathrin-mediated endocytosis was substantially reduced. While in vitro growth, cell morphology, and vacuoles appeared normal, the mutant was impaired in actin patch formation, filamentous growth, biofilm formation, cell wall integrity, and extracellular protease secretion. In addition, susceptibility to various antifungal agents was altered. Consistent with the inability to form hyphae, in an in vitro keratinocyte infection model, the null mutant displayed reduced damage of mammalian adhesion zippers and host cell death. Thus, C. albicans END3 has a role in efficient endocytosis that is required for cell wall integrity, protein secretion, hyphal formation, and virulence-related processes. These findings suggest that impaired endocytosis subsequently affects other secretory pathways, providing evidence of the interconnection between these processes. IMPORTANCE Candida albicans is a fungal commensal organism that can cause serious opportunistic infections in immunocompromised patients leading to substantial complications and mortality. A better understanding of the microbe's biology to develop more effective therapeutic and diagnostic tools is required as invasive candidiasis is a problem of continued clinical importance. This study focuses on endocytosis, an important but incompletely understood cellular mechanism needed to uptake nutrients and communicate with a cell's environment. In this study, we have assessed the role of endocytosis in cell wall integrity, biofilm formation, and tissue invasion in C. albicans. These findings will improve our understanding of cellular mechanisms underlying endocytosis and will inform us of the interconnection with other intracellular transport processes.


Assuntos
Candida albicans , Proteínas Fúngicas , Animais , Parede Celular/metabolismo , Clatrina/metabolismo , Endocitose , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Humanos , Hifas , Mamíferos/metabolismo
13.
Sci Rep ; 12(1): 3407, 2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-35232970

RESUMO

Nutrient foraging by fungi weathers rocks by mechanical and biochemical processes. Distinguishing fungal-driven transformation from abiotic mechanisms in soil remains a challenge due to complexities within natural field environments. We examined the role of fungal hyphae in the incipient weathering of granulated basalt from a three-year field experiment in a mixed hardwood-pine forest (S. Carolina) to identify alteration at the nanometer to micron scales based on microscopy-tomography analyses. Investigations of fungal-grain contacts revealed (i) a hypha-biofilm-basaltic glass interface coinciding with titanomagnetite inclusions exposed on the grain surface and embedded in the glass matrix and (ii) native dendritic and subhedral titanomagnetite inclusions in the upper 1-2 µm of the grain surface that spanned the length of the fungal-grain interface. We provide evidence of submicron basaltic glass dissolution occurring at a fungal-grain contact in a soil field setting. An example of how fungal-mediated weathering can be distinguished from abiotic mechanisms in the field was demonstrated by observing hyphal selective occupation and hydrolysis of glass-titanomagnetite surfaces. We hypothesize that the fungi were drawn to basaltic glass-titanomagnetite boundaries given that titanomagnetite exposed on or very near grain surfaces represents a source of iron to microbes. Furthermore, glass is energetically favorable to weathering in the presence of titanomagnetite. Our observations demonstrate that fungi interact with and transform basaltic substrates over a three-year time scale in field environments, which is central to understanding the rates and pathways of biogeochemical reactions related to nuclear waste disposal, geologic carbon storage, nutrient cycling, cultural artifact preservation, and soil-formation processes.


Assuntos
Hifas , Silicatos , Florestas , Hifas/metabolismo , Silicatos/metabolismo , Solo
14.
Microbiol Res ; 258: 126996, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35247799

RESUMO

Candida albicans is a commensal microorganism of the human microbiota that can be associated with superficial to disseminated infections. This species possesses several attributes that contribute to pathogenesis and virulence, such as the ability to transition from yeast to hyphae forms. During this transition, several changes occur in the fungal cell wall, which is the first point of contact between the pathogen and the host. The cell wall is a bi-layered structure, containing chitin, glucan, and proteins, the latter of which play important roles in pathogenesis. Given the importance of this structure, particularly in filamentation, this review focuses on the studies of C. albicans mutants for genes that encode cell wall-associated proteins that have an important role in the virulence, and also have a role in hyphal morphogenesis. Thus, we highlight some proteins whose mutation is associated with attenuated virulence in vivo and have defective filamentation. We also provide examples of proteins whose inactivation does not impair the filamentation yet are still important for C. albicans virulence.


Assuntos
Candida albicans , Candidíase , Candida albicans/metabolismo , Candidíase/microbiologia , Parede Celular/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Humanos , Hifas/genética , Hifas/metabolismo , Virulência
15.
Int J Mol Sci ; 23(5)2022 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-35269898

RESUMO

Despite abundant research in the field of antifungal drug discovery, fungal infections remain a significant healthcare burden. There is an emerging need for the development of novel antifungals since those currently available are limited and do not completely provide safe and secure protection. Since the current knowledge regarding the physiology of fungal cells and the infection mechanisms is greater than ever, we have the opportunity to use this for the development of novel generations of antifungals. In this review, we selected and summarized recent studies describing agents employing different antifungal mechanisms. These mechanisms include interference with fungal resistance, including impact on the efflux pumps and heat shock protein 90. Additionally, interference with virulence factors, such as biofilms and hyphae; the impact on fungal enzymes, metabolism, mitochondria, and cell wall; and antifungal vaccines are explored. The agents investigated belong to different classes of natural or synthetic molecules with significant attention given also to plant extracts. The efficacy of these antifungals has been studied mainly in vitro with some in vivo, and clinical studies are needed. Nevertheless, there is a large quantity of products employing novel antifungal mechanisms that can be further explored for the development of new generation of antifungals.


Assuntos
Antifúngicos , Micoses , Antifúngicos/metabolismo , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Biofilmes , Parede Celular/metabolismo , Farmacorresistência Fúngica , Humanos , Hifas , Micoses/microbiologia
16.
J Microbiol ; 60(4): 438-443, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35286605

RESUMO

Systemic candidiasis, which is mainly caused by Candida albicans, is a serious acute fungal infection in the clinical setting. In a previous study, we reported that compound 22h (designated as AB-22 in this study), a vinyl sulfate compound, is a fast-acting fungicidal agent against a broad spectrum of fungal pathogens. In this study, we aimed to further analyze the in vitro and in vivo efficacy of AB-22 against filamentation, biofilm formation, and virulence of C. albicans. Under in vitro hyphal growth-inducing condition, AB-22 effectively inhibited germ tube formation and hyphal growth, which are required for the initiation of biofilm formation. Indeed, AB-22 significantly suppressed C. albicans biofilm formation in a dose-dependent manner. Moreover, AB-22 treatment inhibited the normal induction of ALS3, HWP1, and ECE1, which are all required for hyphal transition in C. albicans. Furthermore, AB-22 treatment increased the survival of mice systemically infected with C. albicans. In conclusion, in addition to its fungicidal activity, AB-22 inhibits filamentation and biofilm formation in C. albicans, which could collectively contribute to its potent in vivo efficacy against systemic candidiasis.


Assuntos
Candida albicans , Candidíase , Animais , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Biofilmes , Candidíase/tratamento farmacológico , Candidíase/microbiologia , Hifas , Camundongos
17.
Trends Microbiol ; 30(6): 519-523, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35346551

RESUMO

Macrophages play a crucial role in fungal clearance during Candida albicans infection. In order to eliminate the intracellular fungi and survive, macrophages need to quickly respond to the dimorphic transition by repairing the phagolysosomal membrane. Also, actin-contractile rings were recently suggested as a powerful weapon to dampen hyphal growth.


Assuntos
Candida albicans , Candidíase , Actinas , Candidíase/microbiologia , Humanos , Hifas , Macrófagos/microbiologia
18.
Biomaterials ; 283: 121420, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35245733

RESUMO

Invasive pulmonary aspergillosis is associated with a high mortality rate and poses a direct threat to immunocompromised patients. Here, we present the invasive aspergillosis-on-chip (IAC) model to investigate Aspergillus fumigatus infection in vitro. The model allows the study of the lateral growth and the invasive behaviour of fungal hyphae from the epithelium into the endothelial cell layer in an alveolus-on-chip model. We established an algorithm-based analysis pipeline for three-dimensional confocal microscopy images to visualize and quantify fungal morphology, including hyphal growth and branching. Human macrophages in the IAC model partially inhibited the growth of the fungus, contributed to the release of proinflammatory cytokines (IL-1, IL-6, TNF) and chemokines (IL-8 and MCP-1) associated with an increased number of invasive hyphae. Similar to in vivo, the application of the fungistatic drug caspofungin limited the fungal growth and resulted in morphological changes of the hyphal tree previously described in other studies. The IAC infection model allows the identification and characterization of cellular infection targets and in vitro testing of antifungal drugs in clinically relevant concentrations. It thus represents a promising tool to broaden the understanding of pathogenicity and pathophysiology of invasive aspergillosis.


Assuntos
Aspergilose , Aspergillus fumigatus , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Aspergilose/tratamento farmacológico , Aspergilose/microbiologia , Caspofungina/farmacologia , Caspofungina/uso terapêutico , Humanos , Hifas
19.
Microbiol Spectr ; 10(2): e0147821, 2022 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-35297651

RESUMO

Candida albicans biofilms are resistant to several clinical antifungal agents. Thus, it is necessary to develop new antibiofilm intervention measures. Pseudolaric acid A (PAA), a diterpenoid mainly derived from the pine bark of Pseudolarix kaempferi, has been reported to have an inhibitory effect on C. albicans. The primary aim of the current study was to investigate the antibiofilm effect of PAA when combined with fluconazole (FLC) and explore the underlying mechanisms. Biofilm activity was assessed by tetrazolium {XTT [2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt]} reduction assays. PAA (4 µg/mL) combined with FLC (0.5 µg/mL) significantly inhibited early, developmental, and mature biofilm formation compared with the effect of PAA or FLC alone (P < 0.05). Furthermore, PAA (4 µg/mL) combined with FLC (0.5 µg/mL) produced a 56% reduction in C. albicans biofilm adhesion. The combination of PAA (4 µg/mL) and FLC (0.5 µg/mL) also performed well in inhibiting yeast-to-hypha transition. Transcriptome analysis using RNA sequencing and quantitative reverse transcription PCR indicated that the PAA-FLC combination treatment produced a strong synergistic inhibitory effect on the expression of genes involved in adhesion (ALS1, ALS4, and ALS2) and yeast-to-hypha transition (ECE1, PRA1, and TEC1). Notably, PAA, rather than FLC, may have a primary role in suppressing the expression of ALS1. In conclusion, these findings demonstrate, for the first time, that the combination of PAA and FLC has an improved antibiofilm effect against the formation of C. albicans biofilms by inhibiting adhesion and yeast-to-hypha transition; this may provide a novel therapeutic strategy for treating C. albicans biofilm-associated infection. IMPORTANCE Biofilms are the primary cause of antibiotic-resistant candida infections associated with medical implants and devices worldwide. Treating biofilm-associated infections is a challenge for clinicians because these infections are intractable and persistent. Candida albicans readily forms extensive biofilms on the surface of medical implants and mucosa. In this study, we demonstrated, for the first time, an inhibitory effect of pseudolaric acid A alone and in combination with fluconazole on C. albicans biofilms. Moreover, pseudolaric acid A in combination with fluconazole exerted an antibiofilm effect through multiple pathways, including inhibition of yeast-to-hypha transition and adhesion. This research not only provides new insights into the synergistic mechanisms of antifungal drug combinations but also brings new possibilities for addressing C. albicans drug resistance.


Assuntos
Candida albicans , Fluconazol , Antifúngicos/farmacologia , Biofilmes , Diterpenos , Sinergismo Farmacológico , Fluconazol/farmacologia , Hifas/genética , Testes de Sensibilidade Microbiana
20.
Microbiol Spectr ; 10(1): e0206321, 2022 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-35107348

RESUMO

Septation in filamentous fungi is a normal part of development, which involves the formation of cross-hyphal bulkheads, typically containing pores, allowing cytoplasmic streaming between compartments. Based on previous findings regarding septa and cell wall stress, we hypothesized that septa are critical for survival during cell wall stress. To test this hypothesis, we used known Aspergillus nidulans septation-deficient mutants (ΔsepH, Δbud3, Δbud4, and Δrho4) and six antifungal compounds. Three of these compounds (micafungin, Congo red, and calcofluor white) are known cell wall stressors which activate the cell wall integrity signaling pathway (CWIS), while the three others (cycloheximide, miconazole, and 2,3-butanedione monoxime) perturb specific cellular processes not explicitly related to the cell wall. Our results show that deficiencies in septation lead to fungi which are more susceptible to cell wall-perturbing compounds but are no more susceptible to other antifungal compounds than a control. This implies that septa play a critical role in surviving cell wall stress. IMPORTANCE The ability to compartmentalize potentially lethal damage via septation appears to provide filamentous fungi with a facile means to tolerate diverse forms of stress. However, it remains unknown whether this mechanism is deployed in response to all forms of stress or is limited to specific perturbations. Our results support the latter possibility by showing that presence of septa promotes survival in response to cell wall damage but plays no apparent role in coping with other unrelated forms of stress. Given that cell wall damage is a primary effect caused by exposure to the echinocandin class of antifungal agents, our results emphasize the important role that septa might play in enabling resistance to these drugs. Accordingly, the inhibition of septum formation could conceivably represent an attractive approach to potentiating the effects of echinocandins and mitigating resistance in human fungal pathogens.


Assuntos
Aspergillus nidulans/crescimento & desenvolvimento , Aspergillus nidulans/fisiologia , Parede Celular/fisiologia , Antifúngicos/farmacologia , Aspergillus nidulans/efeitos dos fármacos , Aspergillus nidulans/genética , Parede Celular/efeitos dos fármacos , Parede Celular/genética , Vermelho Congo/farmacologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Hifas/efeitos dos fármacos , Hifas/genética , Hifas/crescimento & desenvolvimento , Hifas/metabolismo , Micafungina/farmacocinética , Viabilidade Microbiana/efeitos dos fármacos , Estresse Fisiológico
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