Search PubMed⌕ Search

Biomedical subjects

T Boekhout

Publications and source records attributed to T Boekhout.

33 records · Page 2Linked to original sources

Molecular typing of Malassezia species with PFGE and RAPD.

The currently recognized seven species of Malassezia all have different karyotypes which do not vary intraspecifically, except in M. furfur which displayed two different karyotypes. In contrast, random amplified polymorphic DNA (RAPD) typing showed the presence of genetic variation in all species. It is concluded that karyotype analysis is useful for species identification, and RAPD typing can be used in epidemiological investigations.

Animals↗

The role of Malassezia species in the ecology of human skin and as pathogens.

The new taxonomic structure of the lipophilic genus Malassezia was presented with key characteristics for the seven described species. Among techniques used for epidemiological surveys, the pulsed field gel electrophoresis (PFGE) was found to be of little value in contrast to randomly amplified polymorphic DNA (RAPD). Immunological studies still yielded conflicting results but at least the immunomodulatory capacity of Malassezia yeasts appeared to be related to the cell wall lipids. A review of Malassezia infections together with the present consensus for their prevention and treatment was also made.

Antibodies, Fungal↗

Molecular characterization of the glyceraldehyde-3-phosphate dehydrogenase gene of Phaffia rhodozyma.

The glyceraldehyde-3-phosphate dehydrogenase (GPD; EC1.2.1.12)-encoding gene (gpd) was isolated from a genomic library of Phaffia rhodozyma CBS 6938. Unlike some other eukaryotic organisms the gpd gene is represented by a single copy in P. rhodozyma. The complete nucleotide sequence of the coding, as well as the flanking non-coding regions was determined. The nucleotide sequence of gpd predicted six introns and a polypeptide chain of 339 amino acids. The codon usage in the gpd gene of P. rhodozyma was highly biased and was significantly different from the codon usage in other yeasts. Phylogenetic analysis of different yeasts and filamentous asco- and basidiomycetes gpd sequences indicated that the gpd gene of P. rhodozyma forms a cluster with the corresponding genes of filamentous basidiomycetes.

Amino Acid Sequence↗

Variability of karyotypes and RAPD types in genetically related strains of Cryptococcus neoformans.

Variation in karyotypes and RAPD patterns of genetically related strains of Cryptococcus neoformans were analyzed. Capsular and filamentous mutants usually differ in their karyotypes from wild-types, but the RAPD patterns were found to be similar. Karyotype differences were observed in most heterothallic matings, but RAPD patterns remained identical. After self-sporulation of a diploid strain, minor chromosomal length polymorphism and minor changes in the RAPD types occurred. Three mechanisms, either alone or in combination, may in varying degrees contribute to the karyotype variation of C. neoformans: (1) mitotically induced changes; (2) karyotype changes as a result of meiotic recombination, and (3) mutagen-induced changes. The present data do not support the meiotic maintenance hypothesis, which claims that the amount of CLP generated is inversely proportional to the frequency of meiosis.

Cryptococcus neoformans↗

Molecular typing of Cryptococcus neoformans: taxonomic and epidemiological aspects.

Pulsed-field gel electrophoresis (PFGE), randomly amplified polymorphic DNA (RAPD) analysis, serotype, and killer toxin sensitivity patterns of a wide range of saprobic, clinical, and veterinary isolates of both varieties of Cryptococcus neoformans were examined. C. neoformans var. neoformans and C. neoformans var. gattii differed in chromosomal makeup, RAPD patterns, and killer sensitivity patterns. These results suggest that there are two separate species rather than two varieties. No clear genetic or phenotypic differences were observed among the clinical, saprobic, and veterinary isolates within each taxon. The serotypes differed substantially in their RAPD characteristics. Geographical clustering was observed among the isolates of C. neoformans var. gattii, but not among the isolates of C. neoformans var. neoformans. The isolates of each taxon that originated from restricted geographical areas often had identical or similar karyotypes and RAPD patterns, suggesting that clonal reproduction had occurred. The combination of PFGE and RAPD analysis allowed us to distinguish almost all isolates. This combination of techniques is recommended for further research on epidemiological, ecological, and population issues.

AIDS-Related Opportunistic Infections↗

Structural and phylogenetic analysis of the actin gene from the yeast Phaffia rhodozyma.

The gene coding for actin from Phaffia rhodozyma was cloned and sequenced. The Phaffia actin gene contains four intervening sequences and the predicted protein consists of 375 amino acids. The structural features of the Phaffia actin introns were studied and compared with actin introns from seven fungi and yeasts with ascomycetous and basidiomycetous affinity. It was shown that the architecture of the Phaffia introns most resembles that of the basidiomycete Filobasidiella neoformans (perfect stage of Cryptococcus neoformans), whereas least resemblance occurs with the ascomycetous yeasts. Based on the intron structure, the ascomycetous yeasts can be accommodated in one group in that their splice site sequences are very similar and show less homology with the other fungi investigated, including Phaffia. It was demonstrated that the Phaffia actin introns cannot be spliced in Saccharomyces cerevisiae, which shows that the differences found in intron structure are significant. Alignment of the Phaffia actin gene with the actin sequences from the yeasts and fungi investigated showed a high level of homology both on the DNA level and on the protein level. Based on these alignments Phaffia showed highest homology with F. neoformans and both organisms were accommodated in the same cluster. In addition, the actin gene comparisons also supported the distant relationship of Phaffia with the ascomycetous yeasts. These results supported the usefulness of actin sequences for phylogenetic studies.

Actins↗

Phylogeny of the yeast genera Hanseniaspora (anamorph Kloeckera), Dekkera (anamorph Brettanomyces), and Eeniella as inferred from partial 26S ribosomal DNA nucleotide sequences.

Partial 26S ribosomal DNA sequences of species assigned to the genera Hanseniaspora, Kloeckera, Dekkera, Brettanomyces, and Eeniella were determined. A phylogenetic analysis of the sequences showed that the genus Eeniella is derived within the genus Brettanomyces and that the genus Hanseniaspora (anamorph Kloeckera) is not closely related to the genus Dekkera (anamorph Brettanomyces). As a consequence, the name Eeniella is reduced to synonymy with the name Brettanomyces. In addition, our data do not support reassignment of certain Hanseniaspora species to the recently revived genus Kloeckeraspora.

Ascomycota↗

Monitoring spread of Malassezia infections in a neonatal intensive care unit by PCR-mediated genetic typing.

Malassezia furfur and Malassezia pachydermatis were isolated from newborn children and incubators in a neonatal intensive care unit. To assess whether persistence or frequent import of the organisms was the cause of the elevated incidence, genetic typing of the strains was performed by PCR-mediated DNA fingerprinting. By using PCR primers aimed at repeat consensus motifs, six different genotypes could be detected in a collection of six M. furfur reference strains. In the case of 10 M. pachydermatis reference strains, nine different genotypes were detected by three different PCR assays. None of these assays could document genetic differences among the clinical isolates of either M. furfur or M. pachydermatis. On the basis of these results it is concluded that within the neonatal intensive care unit the longitudinal persistence of both an M. furfur and an M. pachydermatis strain has occurred and that Malassezia species can persist on incubator surfaces for prolonged periods of time. It can be concluded that PCR fingerprinting is a Malassezia typing procedure that is to be preferred over the analysis of chromosomal polymorphisms by pulsed-field gel electrophoresis in this genus.

Base Sequence↗

The use of karyotyping in the systematics of yeasts.

The use of electrophoretic karyotyping in systematics of yeasts is discussed. New data are provided on the karyotypes of the medically important fungi Hortaea werneckii, Filobasidiella (= Cryptococcus) neoformans, and Malassezia species. Hortaea werneckii has twelve to eighteen bands of chromosomal DNA, ranging in size between 500 and 2300 kb. The karyotypes of Filobasidiella neoformans consist of seven to fourteen bands of chromosomal DNA. The varieties neoformans and bacillispora cannot be separated by their karyotypes, and no obvious correlation was found with serotypes, geography or habitat. All strains of Malassezia pachydermatis studied have similar karyotypes consisting of five bands, whereas in M. furfur, four different karyotypes are prevalent. However, each of these karyotypes is stable.

Chromosomes, Fungal↗

Dimorphism in Itersonilia perplexans: yeast and hyphal phases differ in their sensitivity to mycocins produced by tremellaceous yeasts.

The monokaryotic yeast phase of the heterobasidiomycete Itersonilia perplexans, unlike the hyphal phase, was found to be sensitive to mycocins produced by killer strains of Cryptococcus humicola, Cr. laurentii, Cystofilobasidium bisporidii and Rhodotorula fujisanense. Both the yeast and hyphal phases wer resistant to mycocins of Cr. podzolicus, Filobasidium capsuligenum, Rhodotorula glutinis, Rh. mucilaginosa, Rh. pallida, Sporidiobolus johnsonii, Sb. pararoseus and Sporobolomyces alborubescens. The different sensitivity patterns of yeast and hyphal phases are probably caused by biochemical differences in the cell walls.

Cell Wall↗

Bulleromyces genus novum (Tremellales), a teleomorph for Bullera alba, and the occurrence of mating in Bullera variabilis.

Mating is observed in Bullera alba and B. variabilis, resulting in the formation of dikaryotic mycelium with clamps, haustorial branches, and lateral and terminal dikaryotic, clavate, lageniform or subglobose cells. These cells develop in B. alba into tremellaceous phragmobasidia. Karyogamy has been observed in young non-divided basidia. Germination of the phragmobasidia occurred by acropetal chains of yeast cells, ballistospores or hyphae. Septal pores are dolipores with parenthesomes made up of U-shaped vesicles (Tremellales type). For the teleomorph of B. alba a new genus, Bulleromyces, is proposed, with only one species, viz. Bulleromyces albus.

Basidiomycota↗

Field-emission scanning electron microscopy of the internal cellular organization of fungi.

Internal viewing of the cellular organization of hyphae by scanning electron microscopy is an alternative to observing sectioned fungal material with a transmission electron microscope. To study cytoplasmic organelles in the hyphal cells of fungi by SEM, colonies were chemically fixed with glutaraldehyde and osmium tetroxide and then immersed in dimethyl sulfoxide. Following this procedure, the colonies were frozen and fractured on a liquid nitrogen-precooled metal block. Next, the fractured samples were macerated in diluted osmium tetroxide to remove the cytoplasmic matrix and subsequently dehydrated by freeze substitution in methanol. After critical point drying, mounting, and sputter coating, fractured cells of several basidiomycetes were imaged with field-emission SEM. This procedure produced clear images of elongated and spherical mitochondria, the nucleus, intravacuolar structures, tubular- and plate-like endoplasmic reticulum, and different types of septal pore caps. This method is a powerful approach for studying the intracellular ultrastructure of fungi by SEM.

Fungi↗

Differential killer toxin sensitivity patterns of varieties of Cryptococcus neoformans.

Ten different killer sensitivity types are distinguished within Cryptococcus neoformans, namely four in var. neoformans and six in var. gattii. All strains of the var. gattii investigated were inhibited by killer toxins of C. laurentii CBS 139, whereas those of the var. neoformans were not. Killer sensitivity patterns are an easy-to-use method to differentiate between the two varieties of the clinically important yeast C. neoformans, and may be of help in epidemiological surveys.

AIDS-Related Opportunistic Infections↗