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

Candida glabrata'da Minyon Koloni Oluşumu ve İlaç Direncinin Analizi

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.

Yazar Spotlight: Antifungal Araştırmalar için İlaca Dirençli Candida'da Minyon Kolonileri İndüklemek İçin Yeni Bir Yaklaşım Olarak Fotodinamik Terapi

Rose Bengal Aracılı Fotodinamik Terapi ile Candida glabrate'de Minyon Kolonilerin İndüksiyonu

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.

Candida glabrata'nın Etidyum Bromür ile İndüklenen Minyon Kolonilerinde Mitokondriyal Fonksiyon Analizi

Candida glabrata'yı kültürlemeye başlamak için, tüm deneysel materyalleri çalışma platformuna yerleştirin. Bir maya pepton dekstroz veya YPD Agar plakasından tek bir C.glabrata kolonisi seçin. Ve 3 mililitre YPD ortamı içeren bir cam test tüpüne aktarın.155 RPM'de çalkalayarak 25 santigrat derecede 14 ila 16 saat inkübe edin. İnkübasyondan sonra, 600 nanometre 0.1'lik bir optik yoğunluk elde etmek için kültürü taze YPD içinde seyreltin. Seyreltilmiş kültürü 25 santigrat derecede altı saat boyunca 155 RPM'de çalkalayarak inkübe edin.YPD'de 600 nanometrede 0.1'de 3 mililitreye kadar bir optik yoğunluğa bir maya süspansiyonu hazırlayın. 30 mikrolitre Etidyum bromür stoğu ekleyin ve maya süspansiyonunu gece boyunca 25 santigrat derecede, 155 RPM'de 45 derecelik bir açıyla inkübe edin. Ertesi gün, kültürün optik yoğunluğunu taze YPD ortamı ile 0.65'e ayarlayın.96 oyuklu bir plakada, on kat seyreltme için oyuk başına 180 mikrolitre PBS'ye 20 mikrolitre maya süspansiyonu ekleyin. YPD Agar plakasının dört çeyreğine her seyreltmeden 20 mikrolitre plaka koyun. Plakayı gece boyunca 37 santigrat derecede inkübe edin.Ertesi gün, beş ila 80 kolonili kadranları seçin ve 20 mikrolitre% 20 trifenil tetrazolyum ekleyin. Ve 20 mikrolitre% 20 trifenil tetrazolyum klorürü doğrudan her koloninin merkezine ekleyin. Plakayı 37 santigrat derecede 30 ila 40 dakika inkübe edin.48 bit tam renkli optik tarayıcı kullanarak plakaları 1200 DPI'da tarayın. DNA birleştirici ajan olan Etidyum bromür, C.glabrata'nın minyon mutantlarını etkili bir şekilde indükler.

Research

JoVE Journal

Biology

Mitochondrial Function Analysis in Ethidium Bromide&#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 ...

Biyoloji

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

Candida glabrata'nın Flukonazol ile İndüklenen Minyon Kolonilerinde Mitokondriyal Fonksiyon Analizi

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

Antifungal Fotodinamik Tedaviye Bağlı Candida glabrata Minyon Kolonilerinde Mitokondriyal Fonksiyon Analizi