Assessing Als3 Peptide-Binding Cavity and Amyloid-Forming Region Contributions to Invasion of Human Oropharyngeal Epithelial Cells.

Although it is widely recognized that disruption of reduces the invasion of germ tubes into mammalian oral epithelial cells, the mechanism of this interaction was unexplored. strains with structurally informed mutations to remove adhesive activity of the peptide-binding cavity (PBC) or aggregative activity mediated by the amyloid-forming region (AFR) were assessed for their ability to invade cultured human oropharyngeal epithelial cells. Initial assays utilized untreated fungal and epithelial cells.

Development and Use of a Monoclonal Antibody Specific for the Cell-Surface Protein Hwp1.

The cell-surface protein Hwp1 functions in adhesion to the host and in biofilm formation. A peptide from the Gln-Pro-rich adhesive domain of Hwp1 was used to raise monoclonal antibody (MAb) 2-E8. MAb 2-E8 specificity for Hwp1 was demonstrated using a isolate and strains that expressed at least one allele.

Development and validation of monoclonal antibodies specific for Candida albicans Als2, Als9-1, and Als9-2.

Fungal agglutinin-like sequence (Als) cell-surface glycoproteins, best characterized in Candida albicans, mediate adhesive and aggregative interactions with host cells, other microbes, and abiotic surfaces. Monoclonal antibodies (MAbs) specific for each C. albicans Als protein are valuable reagents for gaining insight into Als protein localization and function.

Deletion Increases the Proportion of Small Cells in a Culture Population: Hypothesizing a Novel Role for Als1.

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 produced a strain that showed delayed germ-tube formation and delayed disease progression in a murine model of disseminated candidiasis. Populations of cultured cells had a higher proportion of smaller cells compared to wild-type or reintegrant control cultures.

Peering Into Pir Protein Function and Comparative Genomics of the Pir Family.

The fungal cell wall, comprised primarily of protein and polymeric carbohydrate, maintains cell structure, provides protection from the environment, and is an important antifungal drug target. Pir proteins (proteins with internal repeats) are linked to cell wall β-1,3-glucan and are best studied in . Sequential deletion of genes produces strains with increasingly notable cell wall damage.

Using Genomics to Shape the Definition of the Agglutinin-Like Sequence () Family in the Saccharomycetales.

The agglutinin-like sequence () family is studied because of its contribution to cell adhesion, fungal colonization, and polymicrobial biofilm formation. The goal of this work was to derive an accurate census and sequence for genes in pathogenic yeasts and other closely related species, while probing the boundaries of the family within the Order Saccharomycetales. Bioinformatic methods were combined with laboratory experimentation to characterize 47 novel loci from 8 fungal species.

Effect of light exposure on growth rate of veterinary clinical dermatophyte isolates.

Background: Veterinary textbooks and literature suggest that exposure to light is inhibitory to growth of clinical dermatophyte isolates.

Hypothesis/objectives: We hypothesized that this idea was derived from experiments that examined the effect of high doses of ultraviolet and visible light exposure on dermatophyte growth, and that exposure to typical room lighting would not adversely affect dermatophyte growth rate.

Methods And Materials: Isolates of common veterinary dermatophytes (three each of Microsporum canis, Nannizia gypsea and Trichophyton benhamiae) were exposed to typical fluorescent room lighting, incubated in a closed drawer, or exposed at close range to fluorescent wide-spectrum light.

Pursuing Advances in DNA Sequencing Technology to Solve a Complex Genomic Jigsaw Puzzle: The Agglutinin-Like Sequence () Genes of .

The agglutinin-like sequence () gene family encodes cell-surface adhesins that interact with host and abiotic surfaces, promoting colonization by opportunistic fungal pathogens such as . Studies of Als protein contribution to adhesion would benefit from an accurate catalog of gene sequences as well as insight into relative gene expression levels. Even in the genomics era, this information has been elusive: genome assemblies are often broken within genes because of their extensive regions of highly conserved, repeated DNA sequences and because there are many similar genes at different chromosomal locations.

Microbial Production of Retinyl Palmitate and Its Application as a Cosmeceutical.

Chemically synthesized retinyl palmitate has been widely used in the cosmetic and biotechnology industry. In this study, we aimed to demonstrate the microbial production of retinyl palmitate and the benefits of microbial retinyl palmitate in skin physiology. A heterologous retinyl palmitate biosynthesis pathway was reconstructed in metabolically engineered using synthetic expression modules from , , and .

Agglutinin-Like Sequence () Genes in the Species Complex: Blurring the Boundaries Between Gene Families That Encode Cell-Wall Proteins.

The agglutinin-like sequence (Als) proteins are best-characterized in and known for their role in adhesion of the fungal cell to host and abiotic surfaces. sequences are often misassembled in whole-genome sequence data because each species has multiple loci that contain similar sequences, most notably tandem copies of highly conserved repeated sequences. The species complex includes , , and , three distinct but closely related species.

SERS immunoassay based on the capture and concentration of antigen-assembled gold nanoparticles.

A simple, rapid, and sensitive immunoassay has been developed based on antigen-mediated aggregation of gold nanoparticles (AuNP) and surface-enhanced Raman spectroscopy (SERS). Central to this platform is the extrinsic Raman label (ERL), which consists of a gold nanoparticle modified with a mixed monolayer of a Raman active molecule and an antibody. ERLs are mixed with sample, and antigen induces the aggregation of the ERLs.

Metabolic engineering of the Stevia rebaudiana ent-kaurene biosynthetic pathway in recombinant Escherichia coli.

The ent-kaurene is a dedicated precursor pool and is responsible for synthesizing natural sweeteners such as steviol glycosides. In this study, to produce ent-kaurene in Escherichia coli, we modularly constructed and expressed two ent-kaurene genes encoding ent-copalyl diphosphate synthase (CPPS) and ent-kaurene synthase (KS) from Stevia rebaudiana known as a typical plant producing steviol glycoside. The CPPS and KS from S.

A proposed mechanism for the interaction between the Candida albicans Als3 adhesin and streptococcal cell wall proteins.

C. albicans binds various bacteria, including the oral commensal Streptococcus gordonii. Published reports documented the role of C.

A piglet model for studying Candida albicans colonization of the human oro-gastrointestinal tract.

Pigs from a variety of sources were surveyed for oro-gastrointestinal (oro-GIT) carriage of Candida albicans. Candida albicans-positive animals were readily located, but we also identified C. albicans-free pigs.

The peptide-binding cavity is essential for Als3-mediated adhesion of Candida albicans to human cells.

The adhesive phenotype of Candida albicans contributes to its ability to colonize the host and cause disease. Als proteins are one of the most widely studied C. albicans virulence attributes; deletion of ALS3 produces the greatest reduction in adhesive function.

Frequent isolation of Arthroderma benhamiae from dogs with dermatophytosis.

Background: Molecular analysis methods have led to many changes in the taxonomy of dermatophyte species.

Hypothesis/objectives: We hypothesized that fungi displaying morphology consistent with a traditional identification of 'Trichophyton mentagrophytes' represent multiple species, consistent with the new taxonomy.

Methods: Fungal specimens (n = 20) were collected directly from animals with dermatophytosis, were among those submitted for diagnostic analysis or were part of historical teaching collections.

A monoclonal antibody specific for Candida albicans Als4 demonstrates overlapping localization of Als family proteins on the fungal cell surface and highlights differences between Als localization in vitro and in vivo.

The Candida albicans agglutinin-like sequence (ALS) family encodes large cell surface glycoproteins that function in adhesion of the fungus to host and abiotic surfaces. Monoclonal antibodies (mAbs) specific for each Als protein were developed to study Als localization on the C. albicans surface.

ALS51, a newly discovered gene in the Candida albicans ALS family, created by intergenic recombination: analysis of the gene and protein, and implications for evolution of microbial gene families.

The Candida albicans ALS family has eight genetic loci, each encoding a large glycoprotein. Als protein function is discussed most frequently in terms of adhesion to host and abiotic surfaces. Analyses of C.

Heterogeneous distribution of Candida albicans cell-surface antigens demonstrated with an Als1-specific monoclonal antibody.

Despite an abundance of data describing expression of genes in the Candida albicans ALS (agglutinin-like sequence) gene family, little is known about the production of Als proteins on individual cells, their spatial localization or stability. Als proteins are most commonly discussed with respect to function in adhesion of C. albicans to host and abiotic surfaces.

Monoclonal antibodies specific for Candida albicans Als3 that immunolabel fungal cells in vitro and in vivo and block adhesion to host surfaces.

Two monoclonal antibodies (MAbs) were raised against the Candida albicans cell-surface glycoprotein Als3 using the N-terminal domain of the protein as the immunogen. ELISA was used to demonstrate the specificity of the MAbs for the Als3 fragment, but not for the corresponding N-terminal domain fragments from other proteins in the Als family. The anti-Als3 MAbs immunolabeled the surface of germ tubes from a diverse collection of wild-type C.

Recognition of Candida albicans Als3 by the germ tube-specific monoclonal antibody 3D9.3.

Monoclonal antibody 3D9.3 (MAb 3D9.3) reacts with the surface of Candida albicans germ tubes and recognizes a protein epitope.

Molecular phylogenetic analysis of a geographically and temporally matched set of Candida albicans isolates from humans and nonmigratory wildlife in central Illinois.

This study explored whether wildlife species serve as the reservoir for human Candida albicans strains in a given geographic area. C. albicans isolates were collected from nonmigratory wildlife admitted to the University of Illinois Wildlife Medical Clinic.

Discovering the secrets of the Candida albicans agglutinin-like sequence (ALS) gene family--a sticky pursuit.

The agglutinin-like sequence (ALS) family of Candida albicans includes eight genes that encode large cell-surface glycoproteins. The high degree of sequence relatedness between the ALS genes and the tremendous allelic variability often present in the same C. albicans strain complicated definition and characterization of the gene family.

Deletion of ALS5, ALS6 or ALS7 increases adhesion of Candida albicans to human vascular endothelial and buccal epithelial cells.

Candida albicans yeast forms deleted for ALS5, ALS6 or ALS7 are more adherent than a relevant control strain to human vascular endothelial cell monolayers and buccal epithelial cells. In the buccal and vaginal reconstituted human epithelium (RHE) disease models, however, mutant and control strains caused a similar degree of tissue destruction. Deletion of ALS5 or ALS6 significantly slowed growth of the mutant strain; this phenotype was not affected by addition of excess uridine to the culture medium.