mitochondrial function analysis in fluconazole&#45;induced petite colonies of candida glabrata

Analyse af dannelse af petite koloni og lægemiddelresistens i Candida glabrata

Fungal infections, particularly those caused by Candida albicans and increasingly drug-resistant Candida glabrata, pose a serious global threat1. These infections can be deadly, especially for hospitalized patients and those with weakened immune systems. Rising antifungal resistance threatens the control of invasive candidiasis, a severe fungal infection with high mortality, especially from Candida albicans2. Resistant strains hinder effective treatment, potentially increasing both complexity and death rates. In Alameda County, California, USA, C. glabrata has become the most prevalent invasive species3. This shift in the prevalence and distribution of Candida species may be influenced by local healthcare practices, patient demographics, the utilization of antifungal agents, and the prevalence of risk factors for Candida infections.
Petite mutants in Candida, lacking functional mitochondria, reveal how this organelle affects drug response, virulence, and stress resistance4,5. C. glabrata readily forms these colonies, gaining sensitivity to polyenes while losing it to azoles6. Azole sensitivity and respiratory function are intricately linked, with diminished respiration leading to resistance via mitochondrial DNA loss7. Petite colonies of C. glabrata with azole resistance have been isolated from human stool samples from a bone marrow transplant recipient undergoing fluconazole treatment8 and from blood culture bottles of patients with bloodstream infections9. Their potential implications in drug resistance, virulence, and stress response highlight their clinical significance. Additionally, their distinct properties make them valuable tools for investigating fundamental questions in mitochondrial biology5. As research into petite mutants continues, their applications in both clinical and basic research are likely to expand.
This study discovered that photodynamic therapy (PDT) can induce petite colonies in C. glabrata, expanding the range of methods beyond the traditional techniques of exposing C. glabrata to ethidium bromide or fluconazole.

Forfatter Spotlight: Fotodynamisk terapi som en ny tilgang til at inducere små kolonier i lægemiddelresistent Candida til svampedræbende forskning

Induktion af små kolonier i Candida-glabrat via rosenbengalmedieret fotodynamisk terapi

The research aims to use photodynamic therapy, also known as PDT, to fight against drug-resistant Candida infection. In this study, we are trying to answer how effectively PDT kills the fungi, and how the virulence of the fungi changes after the treatment. We found petite colonies from the after PDT.Present methods for inducing petite colonies rely on ethidium bromide or fluconazole, which may work through different pathways on drug resistance and the stress responses, compared to PDT. Recently, petite colonies of Candida have become a significant concern in the clinic because they may resist medication and increase mortality rates. Photodynamic therapy has become one of the antifungal treatments and our finding helps breach the gap between traditional methods used to study petite colonies.Our findings provide evidence that PDT can induce petite colonies in drug resistant Candida in-vitro, presenting a groundbreaking method to study petite colonies. We'll definitely delve deeper into this field. There are many questions to be answered.How and why do petite colonies form after PDT stress? Do different Candida species respond differently? I believe it's a never ending story to be explored.

Mitokondriefunktionsanalyse i fluconazol-inducerede petite kolonier af Candida glabrata

Til at begynde med skal du placere Candida glabrata-kulturen og YPD-mediet på en arbejdsplatform. Juster den optiske tæthed af gærcellesuspension ved 600 nanometer til 0,1 med YPD-medium. Overfør en milliliter af cellesuspensionen til et rør på fem milliliter.Dyp en steril vatpind i gærcellesuspensionen. Vab bomulden frem og tilbage på Mueller Hinton Agar-pladen, og drej to gange ved 60 grader. Drej derefter omkredsen for jævn dækning.Opdel pladen i tre lige store sektorer ved hjælp af en markør. Steriliser tangen over en flamme i et til to sekunder. Efter afkøling skal du placere en tom disk i midten af hver sektor.Tilsæt 12,5 mikroliter fluconazolfond til hver disk. Bland grundigt for at undgå bobler, og inkuber ved 37 grader Celsius i 20 til 24 timer. Den følgende dag tilsættes 20 mikroliter 20% Triphenyltetrazoliumchlorid direkte på midten af hver koloni.Inkuber pladen ved 37 grader Celsius i 30 til 40 minutter. Scan pladen ved 1200 DPI ved hjælp af en 48-bit optisk farvescanner. Fluconazol-behandlingen inducerer effektivt små mutanter af C. glabrata.En klar cirkulær zone uden vækst omgav fluconazolskiven i T-nulkulturen, hvilket indikerer mindre lægemiddelresistens i forældrenes svampe. I T-tre-kulturen dannede mange små kolonier nær disken, hvilket tyder på lægemiddelresistens efter tre gentagne fotodynamiske terapibehandlinger.

Research

JoVE Journal

Biology

Mitochondrial Function Analysis in Fluconazole&#45;Induced Petite Colonies of Candida glabrata

Induction of Petite Colonies in Candida glabrate via Rose Bengal-Mediated Photodynamic Therapy

Facing a 40% mortality rate in candidemia patients, drug-resistant Candida and their petite mutants remain a major treatment challenge. Antimicrobial ...

Mitochondrial Function Analysis in Ethidium Bromide&#45;Induced Petite Colonies of Candida glabrata

Mitokondriefunktionsanalyse i ethidiumbromidinducerede petitekolonier af Candida glabrata

Mitochondrial Function Analysis in Antifungal Photodynamic Therapy Induced Petite Colonies of Candida glabrata

Mitokondriefunktionsanalyse i svampedræbende fotodynamisk terapi inducerede petite kolonier af Candida glabrata