'Evaluation of fungal biofilm formation can be performed using several techniques. In this protocol, we describe methods used to form Candida biofilms on three different medical device substrates (denture strips, catheter disks and contact lenses) to quantify them and to evaluate their architecture and drug susceptibility. Biofilm formation involves adhesion ... More
Mechanistic aspects in the generation of apparent ultrahigh efficiencies for colloidal (microbial) electrokinetic separations.
AuthorsArmstrong DW, Girod M, He L, Rodriguez MA, Wei W, Zheng J, Yeung ES
JournalAnal Chem
PubMed ID12433083
'Under specific experimental conditions, the electrokinetic separation of certain microorganisms can produce peaks of very high apparent efficiencies (approximately 10(6)-10(10) theoretical plates/m). This is unusual in that no deliberate focusing mechanism was employed. To investigate this process further, the separation was monitored in real time using a charge-coupled device (CCD) ... More
Pleiotropic effects of HIV-1 protein R (Vpr) on morphogenesis and cell survival in fission yeast and antagonism by pentoxifylline.
AuthorsZhao Y, Yu M, Chen M, Elder RT, Yamamoto A, Cao J
JournalVirology
PubMed ID9657945
'Expression of HIV-1 Vpr causes cell cycle G2 arrest, change in cell shape, and cell death over a large evolutionary distance ranging from human to yeast cells. As a step toward understanding these highly conserved Vpr functions, we have examined the effect of Vpr on cytoskeletal elements and the viability ... More
Mechanism of fluconazole resistance in Candida albicans biofilms: phase-specific role of efflux pumps and membrane sterols.
AuthorsMukherjee PK, Chandra J, Kuhn DM, Ghannoum MA
JournalInfect Immun
PubMed ID12874310
'Candida albicans biofilms are formed through three distinct developmental phases and are associated with high fluconazole (FLU) resistance. In the present study, we used a set of isogenic Candida strains lacking one or more of the drug efflux pumps Cdr1p, Cdr2p, and Mdr1p to determine their role in FLU resistance ... More
Adaptation of FUN-1 and Calcofluor white stains to assess the ability of viable and nonviable yeast to adhere to and be internalized by cultured mammalian cells.
AuthorsHenry-Stanley MJ, Garni RM, Wells CL,
JournalJ Microbiol Methods
PubMed ID15369865
'The FUN-1 and Calcofluor white stains can be used in concert to assess the ability of viable and nonviable yeast to adhere to, and be internalized by, host mammalian cells in vitro. With this method, only extracellular yeast stain with Calcofluor, dead yeast cells have diffuse cytoplasmic yellow-green fluorescence, and ... More
Rapid susceptibility testing of fungi by flow cytometry using vital staining.
AuthorsWenisch C, Linnau KF, Parschalk B, Zedtwitz-Liebenstein K, Georgopoulos A
JournalJ Clin Microbiol
PubMed ID8968873
'A 1-h assay for antifungal susceptibility testing measuring the impairment of fungal metabolic activity was developed. Yeast viability was analyzed by flow cytometry with a novel fluorescent probe, FUN-1, which emits a red fluorescence when the yeast is metabolically active. For nine Candida albicans strains tested, this method yielded results ... More
Determination of product inhibition of CBH1, CBH2, and EG1 using a novel cellulase activity assay.
AuthorsDu F, Wolger E, Wallace L, Liu A, Kaper T, Kelemen B,
JournalAppl Biochem Biotechnol
PubMed ID19830597
The hydrolysis of lignocellulosic biomass by degrading enzymes (cellulases) has emerged as a promising process within the bio-ethanol industry. Yet, understanding all the intricacies of how these enzymes work has been a challenging task. Substrate-enzyme interaction in complex feed mixtures, the recalcitrance of the crystalline structure of cellulose and enzyme ... More
MMM1 encodes a mitochondrial outer membrane protein essential for establishing and maintaining the structure of yeast mitochondria.
AuthorsBurgess SM, Delannoy M, Jensen RE
JournalJ Cell Biol
PubMed ID8089172
In the yeast Saccharomyces cerevisiae, mitochondria are elongated organelles which form a reticulum around the cell periphery. To determine the mechanism by which mitochondrial shape is established and maintained, we screened yeast mutants for those defective in mitochondrial morphology. One of these mutants, mmm1, is temperature-sensitive for the external shape ... More
Development of the FUN-1 family of fluorescent probes for vacuole labeling and viability testing of yeasts.
AuthorsMillard PJ, Roth BL, Thi HP, Yue ST, Haugland RP
JournalAppl Environ Microbiol
PubMed ID9212436
A new family of fluorescent probes has been developed for assessing the viability and metabolic activity of yeasts. This class of halogenated unsymmetric cyanine dyes is exemplified by the FUN-1 [2-chloro-4-(2,3-dihydro-3-methyl-(benzo-1,3-thiazol-2-yl)- methylidene)-1-phenylquinolinium iodide] stain, a membrane-permeant nucleic acid-binding dye that has been found to give rise to cylindrical intravacuolar structures ... More
Comparison of biofilms formed by Candida albicans and Candida parapsilosis on bioprosthetic surfaces.
AuthorsKuhn DM, Chandra J, Mukherjee PK, Ghannoum MA
JournalInfect Immun
PubMed ID11796623
Little is known about fungal biofilms, which may cause infection and antibiotic resistance. In this study, biofilm formation by different Candida species, particularly Candida albicans and C. parapsilosis, was evaluated by using a clinically relevant model of Candida biofilm on medical devices. Candida biofilms were allowed to form on silicone ... More
Unveiling Cells' Local Environment during Cryopreservation by Correlative In Situ Spatial and Thermal Analyses.
Authors
JournalJ Phys Chem Lett
PubMed ID32841035
Caloric restriction extends yeast chronological lifespan via a mechanism linking cellular aging to cell cycle regulation, maintenance of a quiescent state, entry into a non-quiescent state and survival in the non-quiescent state.