Ritu Pasrija

This book is a compilation of protocols for conducting experiments with a variety of yeasts, including S. cerevisiae, Candida albicans, non-albicans Candida sp., Cryptococcus sp.; and molds, including Aspergillus sp., Fusarium sp., Mucor, Rhizopus. The chapters not only discuss National Committee for Clinical Laboratory Standards (NCCLS) accredited protocols but also emphasize the significance of Biosafety Level-2 (BSL-2) facilities. Subsequent chapters cover the essentials of handling, culturing, preserving, and reviving various fungal pathogens. Moreover, advanced techniques are explored, including counting colony-forming units (CFU), growth curve preparation, and microtiter plate assays. A separate chapter discusses the protocols for disc-diffusion, spot and kill-kinetics assays, addressing analyses like a checkerboard and Fractional Inhibitory Concentration (FIC) index assessments. Additionally, the book covers the formation of biofilm, capsule and melanin, fungal cell adhesion, and sterols extraction. Finally, it elaborates on the optimized impact achieved by assessing mitochondrial fitness through non-fermentable carbon. This book is relevant for mycologists and scientists working in the field, providing insights into fungal pathogens and protocols for handling different fungal species.

This book is a compilation of protocols for conducting experiments with a variety of yeasts, including S. cerevisiae, Candida albicans, non-albicans Candida sp., Cryptococcus sp.; and molds, including Aspergillus sp., Fusarium sp., Mucor, Rhizopus. The chapters not only discuss National Committee for Clinical Laboratory Standards (NCCLS) accredited protocols but also emphasize the significance of Biosafety Level-2 (BSL-2) facilities. Subsequent chapters cover the essentials of handling, culturing, preserving, and reviving various fungal pathogens. Moreover, advanced techniques are explored, including counting colony-forming units (CFU), growth curve preparation, and microtiter plate assays. A separate chapter discusses the protocols for disc-diffusion, spot and kill-kinetics assays, addressing analyses like a checkerboard and Fractional Inhibitory Concentration (FIC) index assessments. Additionally, the book covers the formation of biofilm, capsule and melanin, fungal cell adhesion, and sterols extraction. Finally, it elaborates on the optimized impact achieved by assessing mitochondrial fitness through non-fermentable carbon. This book is relevant for mycologists and scientists working in the field, providing insights into fungal pathogens and protocols for handling different fungal species.

La Candida albicans un fungo opportunistico diploide che causa infezioni in pazienti immunocompromessi e debilitati. L'ampia diffusione e l'uso prolungato degli azoli negli ultimi anni ha portato al rapido sviluppo del fenomeno della resistenza agli azoli, che rappresenta una grave minaccia per la terapia antimicotica. Vari meccanismi contribuiscono allo sviluppo della resistenza nella Candida, come la sovraespressione di pompe codificate da geni appartenenti alle superfamiglie di trasportatori ATP-Binding Cassette (ABC) e Major Facilitator (MFS), e/o mutazioni nell'enzima bersaglio degli azoli, ecc. Sulla base di numerosi studi, stata individuata una stretta interazione tra i lipidi di membrana e le proteine della pompa di estrusione dei farmaci. stato dimostrato che l'azione degli antimicotici modulata da sottili modifiche della composizione dei lipidi di membrana.

A Candida albicans um fungo diploide oportunista que causa infec es em doentes imunocomprometidos e debilitados. A utiliza o generalizada e prolongada de az is nos ltimos anos levou ao r pido desenvolvimento do fen meno da resist ncia aos az is, que representa uma grande amea a para a terapia antif ngica. V rios mecanismos contribuem para o desenvolvimento da resist ncia em Candida, como a sobreexpress o de bombas codificadas por genes pertencentes s superfam lias de transportadores ATP-Binding Cassette (ABC) e Major Facilitator Superfamilies (MFS), e/ou muta es na enzima-alvo dos az is, etc. Com base em v rios estudos, verificou-se uma intera o estreita entre os l pidos da membrana e as prote nas da bomba de extrus o de f rmacos. Foi demonstrado que a a o dos antif ngicos modulada por uma altera o subtil da composi o lip dica da membrana.

Candida albicans est un champignon diplo de opportuniste qui provoque des infections chez les patients immunod prim s et affaiblis. L'utilisation tendue et prolong e des azoles au cours des derni res ann es a conduit au d veloppement rapide du ph nom ne de r sistance aux azoles qui constitue une menace majeure pour la th rapie antifongique. Divers m canismes contribuent au d veloppement de la r sistance chez Candida, tels que la surexpression des pompes cod es par des g nes appartenant la superfamille des transporteurs ABC (ATP-Binding Cassette) et MFS (Major Facilitator Superfamilies), et/ou des mutations dans l'enzyme cible des azoles, etc. Plusieurs tudes ont mis en vidence une interaction troite entre les lipides membranaires et les prot ines des pompes d'extrusion des m dicaments. Il a t d montr que l'action des antifongiques est modul e par une modification subtile de la composition des lipides membranaires.

Candida albicans ist ein opportunistischer diploider Pilz, der bei immungeschw chten und geschw chten Patienten Infektionen verursacht. Die weite Verbreitung und der anhaltende Einsatz von Azolen in den letzten Jahren hat zur raschen Entwicklung des Ph nomens der Azolresistenz gef hrt, die eine gro e Bedrohung f r die antimykotische Therapie darstellt. Verschiedene Mechanismen tragen zur Entwicklung einer Resistenz bei Candida bei, wie z. B. die berexpression von Pumpen, die von Genen kodiert werden, die zu den ATP-Bindungskassetten (ABC) und den Hauptf rderer-Superfamilien von Transportern (MFS) geh ren, und/oder Mutationen im Zielenzym von Azolen usw. Basierend auf mehreren Studien wurde eine enge Wechselwirkung zwischen Membranlipiden und Arzneimittel-Extrusionspumpenproteinen festgestellt. Es wurde nachgewiesen, dass die Wirkung von Antimykotika durch eine geringf gige nderung der Membranlipidzusammensetzung moduliert wird.

Candida albicans is an opportunistic diploid fungus that causes infection in immunocompromised and debilitated patients. Wide spread and prolonged usage of azoles in recent years has led to the rapid development of the phenomenon of azole resistance which poses a major threat to antifungal therapy. Various mechanisms contribute towards the development of resistance in Candida such as overexpression of pumps encoded by genes belonging to ATP-Binding Cassette (ABC) and Major Facilitator Superfamilies of transporters (MFS), and/or mutations in the target enzyme of azoles etc. Based on several studies, a close interaction between membrane lipids and drug extrusion pump proteins has been realized. It has been shown that the action of antifungals is modulated by subtle modification of the membrane lipid composition.