Esporotricosis en Abancay, Perú / Sporotrichosis in Abancay, Perú

En la ciudad de Abancay se realizó un estudio con la finalidad de conocer algunas de las características de la esporotricosis. Se efectuó una encuesta clínico-epidemiológica, con toma de muestras de lesiones para determinar la presencia de Sporothrix schenckii mediante cultivo, en 39 pacientes sospechosos de esporotricosis que acudieron a consulta externa del Centro Médico Santa Teresa, entre enero y octubre de 1986. Los criterios de inclusión fueron: características de lesiones, duración de la enfermedad y falta de respuesta a otros tratamientos. Se encontró que en esta ciudad la esporotricosis se presenta más frecuentemente en los menores de 15 años; la mayor parte presenta una lesión única y generalmente en la cara; la variedad de lesión más frecuente fue la cutánea fija. Se concluye que Abancay es una zona endémica de esporotricosis; sin embargo falta estudiar más esta micosis para establecer su mecanismo de transmisión y el nicho ecológico, y así poder determinar las medidas de control más adecuadas.

The CHL 1 (CTF 1) gene product of Saccharomyces cerevisiae is important for chromosome transmission and normal cell cycle progression in G2/M.

We have analyzed the CTF1 gene, identified in a screen for mutants with decreased chromosome transmission fidelity and shown to correspond to the previously identified chl1 mutation. Chl1 null mutants exhibited a 200-fold increase in the rate of chromosome III missegregation per cell division, and near wild-type rates of marker homozygosis on this chromosome by mitotic recombination. Analysis of the segregation of a marker chromosome indicated that sister chromatid loss (1:0 segregation) and sister chromatid non-disjunction (2:0 segregation) contributed equally to chromosome missegregation. A genomic clone of CHL1 was isolated and used to map its physical position on chromosome XVI. Nucleotide sequence analysis of CHL1 revealed a 2.6 kb open reading frame with a 99 kd predicted protein sequence that contained two PEST sequences and was 23% identical to the coding region of a nucleotide excision repair gene, RAD3. Domains of homology between these two predicted protein sequences included a helix-turn-helix motif and an ATP binding site containing a helicase consensus. Mutants lacking the CHL1 gene product are viable and display two striking, and perhaps interrelated, phenotypes: extreme chromosome instability and a delay in cell cycle progression in G2/M. This delay is independent of the cell cycle checkpoint that requires the function of the RAD9 gene.

Characterization of a DL-dityrosine-containing macromolecule from yeast ascospore walls.

We have shown previously that the outer layers of yeast ascospore walls contain dityrosine and that this amino acid is a major component of the cross-linked peptides present in the spore wall (Briza, P., Winkler, G., Kalchhauser, H., and Breitenbach, M. (1986) J. Biol. Chem. 261, 4288-4294). We now present evidence that dityrosine is located in the outermost layer and that it is in the DL-configuration. Although the proteins (peptides) of the spore wall are insoluble, the macromolecule containing dityrosine can be solubilized by partial acid hydrolysis of spore walls. Analysis of this macromolecule indicates that it contains more than 50 mol% dityrosine and a very limited number of other amino acids. Interestingly, part of the dityrosine of spore walls is present in the DL-configuration. We speculate that not only the high degree of cross-links in the outermost layer but also the D-configuration of part of the alpha-C-atoms of dityrosine could contribute to the spores' resistance to lytic enzymes.

Analysis of Pneumocystis carinii cyst wall. II. Sugar composition.

Pneumocystis carinii cysts are capable of resisting host defenses and antimicrobial drugs and are therefore thought to be responsible for relapses of P. carinii pneumonia in AIDS and other immunocompromised patients. The interaction of P. carinii with its host, and other P. carinii, might be mediated by molecules which form the outer surfaces of this organism. Carbohydrates are known to play many roles in cell-cell adhesion, and have been detected on the surface of P. carinii by lectin labeling experiments. In this study P. carinii cyst wall material was obtained from Zymolyase treatment. Alditol acetate derivatives of neutral and amino sugars or trimethylsilyl derivatives of methyl glycosides were prepared from the monosaccharides released from the sample by acid hydrolysis. Analyses were done by a combination of gas chromatography and mass spectrometry. Glucose was found to be the major sugar constituent. Mannose and galactose were present in equal ratios. A lesser amount of N-acetyl-D-glucosamine, and trace amounts of ribose and sialic acid were present in the cyst wall samples analyzed. These sugars may mediate P. carinii-host interaction and play an important protective role by creating a permeability barrier around the cyst.

Evaluation of methods for enumerating microorganisms in filter samples from highly contaminated occupational environments.

Scanning electron microscopy (SEM), light microscopy (LM), epifluorescence microscopy (FM), and culture were used to assess catches of microorganisms in parallel air samples on membrane filters from heavily contaminated working environments that differed in the relative abundance of bacteria, actinomycetes, and fungal spores. Except in pig houses, estimates by SEM and LM were similar, but those by FM and culture were smaller. However, in pig houses, the fluorescent stain enabled bacteria on skin scales, not seen by SEM or LM, to be counted. Although counts obtained by culturing were always smaller than those obtained by SEM or LM, they sometimes exceeded those obtained by FM. Counts suggested that 0.1-68% of bacteria + actinomycetes and 3-98% of fungal spores were viable. However, samples for culturing may have contained larger aggregates than parallel samples collected within a sampling apparatus. All spore types recognized by LM included aggregates--those of bacteria + actinomycetes sometimes exceeding 200 units, while Wallemia sebi spore aggregates were never larger than 3 spores. The size distributions of all types approximated to log-normal, although single spores and small aggregates of bacteria + actinomycetes were perhaps underrepresented. When spores were counted directly on the filter surface, as by SEM and LM, allowance was necessary for heavier deposition of particles near the center of filters by distributing counting fields systematically over the whole filter or a sector of it. Deposition was more uniform in graphite-filled polypropylene filter holders used open-faced. Losses within filter holders and during transportation from sampling site to laboratory were small. The precision of counting spore-containing particles by LM and SEM was better than that of counting individual spores. No such difference was found for FM because many large spore-containing particles were dispersed during preparation.

Molecular cloning of the gene for the E1 alpha subunit of the pyruvate dehydrogenase complex from Saccharomyces cerevisiae.

The E1 alpha and E1 beta subunits of the pyruvate dehydrogenase complex from the yeast Saccharomyces cerevisiae were purified. Antibodies raised against these subunits were used to clone the corresponding genes from a genomic yeast DNA library in the expression vector lambda gt11. The gene encoding the E1 alpha subunit was unique and localized on a 1.7-kb HindIII fragment from chromosome V. The identify of the gene was confirmed in two ways. (a) Expression of the gene in Escherichia coli produced a protein that reacted with the anti-E1 alpha serum. (b) Gene replacement at the 1.7-kb HindIII fragment abolished both pyruvate dehydrogenase activity and the production of proteins reacting with anti-E1 alpha serum in haploid cells. In addition, the 1.7-kb HindIII fragment hybridized to a set of oligonucleotides derived from amino acid sequences from the N-terminal and central regions of the human E1 alpha peptide. We propose to call the gene encoding the E1 alpha subunit of the yeast pyruvate dehydrogenase complex PDA1. Screening of the lambda gt11 library using the anti-E1 beta serum resulted in the reisolation of the RAP1 gene, which was located on chromosome XIV.

Ubiquitin immunoreactivity shows several proteins varying with development and sporulation in the basidiomycete Coprinus cinereus.

Crude extracts of mycelia and basidiocarp primordia in the basidiomycete Coprinus cinereus were resolved on sodium dodecyl sulfate--polyacrylamide gels, and ubiquitin and several proteins were detected by immunoblotting with anti-ubiquitin antibody. The molecular masses of the proteins detected were 30,900, 28,600, 27,800, 26,300, 22,500, and 15,400 daltons, respectively. Relative levels of ubiquitin and most of the ubiquitin-immunoreactive proteins in basidiocarp primordium formation increased and in basidiocarp maturation decreased in cap and upper stipe, while in lower stipe became high except for the 27,800 dalton protein and ubiquitin. During sporulation, ubiquitin and all the ubiquitin-immunoreactive proteins tended to decrease in the cap of the young wild-type basidiocarp. The levels of 30,900 and 15,400 dalton proteins increased transiently at 6-10 h after the beginning of the last light period, while ubiquitin decreased markedly. No correlation was observed between changes in levels of the ubiquitin-immunoreactive proteins and the blocked stages in sporulation-deficient mutants.

[Fungal spores in the atmosphere at Besançon, France: seasonal and annual variations during 1988 and 1989]. / Les spores fungiques dans l'atmosphère de Besançon, France: variations saisonnières et annuelles au cours des années 1988 et 1989.

Airborne spora in the atmosphere of Besançon were weekly determined, searching links between appearance of species and seasonal allergies. By comparison of the results of the two years, it is easy to see many variations. At first, the number of colonies collected is very different, 2,990 colonies in 1988, 9,841 in 1989. During 1989, some genera are present in greatest quantities. It is the case of Cladosporium and Epicoccum in summer and Aspergillus in autumn. For the allergologists, Alternaria, very abundant during 1989, is perhaps the outstanding incident, this genus being considerated as very allergenic.

Fungi carried from farmers' work into farm homes.

Airborne fungal spore concentrations and main fungal genera were compared in rural and urban living environments in Finland during the winter. In addition to conventional viable fungal spore counts (based on the six-stage impactor sampling and cultivation), total spore concentrations were obtained by scanning electron microscope (SEM) investigation of filter samples. The viable spore counts were only 0.2%-25% of the number of total spore aggregates. A high correlation between these two methods was noted, however, at the recommended measuring ranges of the methods. In the farm houses, viable and total spore levels were 10(3) to 10(4) colony forming units/m3 (cfu/m3) and 10(4) to 10(5), spores/m3, respectively. These counts were 10-10(3)-fold higher than the concentrations in an urban apartment. The spore levels of farmers' homes, however, were somewhat lower than those observed in their cow barns. Aspergillus, Cladosporium, and Penicillium spores were present in both urban and rural environments. Actinomycetes and some fungal genera--such as Acremonium, Alternaria, Botrytis, and Chrysosporium--which were detected in cow barns and in farm houses, were not present in urban environment. The results indicated that airborne fungal spores may be carried from cow barns to farmers' homes.

Characterization of exposure to molds and actinomycetes in agricultural dusts by scanning electron microscopy, fluorescence microscopy and the culture method.

Air samples from 79 farms with 10(5) to 10(11) microorganisms/m3 were analyzed by scanning electron microscopy (SEM), fluorescence microscopy (FM), and the culture method. The total exposure to microorganisms (particularly actinomycetes) was underestimated when assessed as colony-forming units (cfu). The average cfu count was one-sixth of the total count according to SEM or FM, and the individual variability was great. This occurrence was partly explained by the aggregation of spores. Single spores accounted for 2-65% of all spores in 35 samples. There was an average of three spores/particle, and 93 (range 67-100)% of the spores were single or in aggregates of respirable size. Aggregation was more pronounced for actinomycetes and at high spore counts. Actinomycetes and bacteria could not be distinguished by FM. Bacteria (other than actinomycetes) were not detected by SEM, yet the total count of microorganisms was similar for FM and SEM. Most particles were spores from actinomycetes and fungi of the genera Aspergillus or Penicillium.

An aerobiological survey of allergens in al Khobar, Saudi Arabia.

A nationwide aerobiologic study is in progress in Saudi Arabia using Burkard 7-day volumetric spore traps to determine the major airborne allergens and their seasonal patterns. Eights months readings have been completed at Al-Khobar, an important coastal city on the Arabian Gulf. Pollen levels showed a double season. An autumnal peak reached its maximum in October rising sharply from the low summer values before falling during the short winter then rising again in springtime. Both local and imported flora were represented with chenopodiaceae, grasses and Ambrosia as the most common botanical groups, identification of the most significant individual species is still in progress. Fungal spores also show distinctive seasonal patterns. In descending rank order from the most common genera were Cladosporium, Ustilago, Alternaria, with Chaetomium and Ulocladium as consistent but minor components. Basiodiospores and Ascospores represented less than 10% of the total spore population, indicative of the dry nature of the climate. Desert dust added an important irritant to the Saudi atmosphere but a major contaminating factor to the aerobiological material being analysed.

Immunoprint analysis of Calvatia cyathiformis allergens. I. Reactivity with individual sera.

Basidiomycete allergens have received scant attention to date, compared to allergens from deuteromycetes. This is true even though in previous studies, 32% of atopic patients with respiratory allergies were skin test reactive to basidiospore extracts. Since sufficient quantities of Calvatia cyathiformis spores were available, their allergens were sequentially fractionated by gel filtration (GF) and hydrophobic interaction chromatography (HIC). Unfractionated extract (crude), GF, and HIC fractions were electrofocused in polyacrylamide gel (pH 3.5 to 9.5) and immunoprinted onto CNBr-activated nitrocellulose (0.2 microns) filters. Sera from 19 skin prick test positive and 10 negative subjects, with comparable total IgE levels, were used to screen individual strips of each blot. Blots were reacted with 125I-labeled anti-IgE and analyzed by autoradiography. IgE from 79%, 89%, and 89% of the positive sera bound to crude extract, GF, and HIC, respectively; IgE from none of the negative sera bound to crude extract. A series of bands (pH 3.6 to 4.6) reacted with 63%, one band (pH 6.6) reacted with 68%, and one band (pH 9.3) reacted with 63% of the sera tested. These studies demonstrate at least three important groups of allergens in C. cyathiformis spores. This characterization allows assignment of initial C. cyathiformis allergen designations and development of a purification protocol.

Respiratory symptoms in damp homes. A pilot study.

The association between living in damp homes and the prevalence of health symptoms was investigated in a population of 519 occupants (adults and children) of 185 homes. Positive associations were found between the reporting of respiratory and some other health symptoms and living in a damp home. The concentration of variable mould spores in indoor air was measured, using modified Andersen samplers, in the living rooms of a sample of 36 homes. The results were compared with the occurrence of dampness characteristics in their homes as reported by the occupants. Homes with at least two dampness characteristics showed higher average spore counts and higher prevalence of respiratory symptoms.

Ultrastructural study of galacturonic acid distribution in some pathogenic fungi using gold-complexed Aplysia depilans gonad lectin.

Aplysia gonad lectin, isolated from the mollusc Aplysia depilans, was successfully conjugated to colloidal gold and used for ultrastructural detection of galacturonic acids in some pathogenic fungi. These sugar residues were found to occur in the fibrillar sheath surrounding hyphal cells of Ascocalyx abietina and in intravacuolar dense inclusions of this fungus spores. In hyphae and spores of Ophiostoma ulmi, galacturonic acids were detected mainly in the outermost wall layers. In contrast, these saccharides appeared associated with the innermost wall layers and especially the plasma membrane of Verticillium albo-atrum cells. Galacturonic acids were found to be absent in cells of Fusarium oxysporum f.sp. radicis-lycopersici and Candida albicans. These cytochemical data indicate therefore that a heterogeneity in wall composition exists between ascomycete fungi. The significance of the presence of galacturonic acids in the cell walls of certain fungi is still open to question.

The major aeroallergens in Guangxi, China.

Over a 2-year period we have identified pollen grains from 48 families of grasses, as well as mould spores and mite particles during air sampling in Guangxi Province. The major aeroallergens were Artemisia, Moraceae and Euophoribiacea, and the spores of Aspergillus, Penicillinum, Cephalosporium and Helminthosporium. Mites were probably also one of the major outdoor aeroallergens. Our investigations also included inspection of the vegetation of the geographical area involved, as well as skin testing on 774 subjects using extracts of 37 aeroallergens. We believe that this work has provided fundamental information on seasonal allergy in Southern China and South-east Asia.

Visualization of exocellular lectins in the entomopathogenic fungus Conidiobolus obscurus.

We demonstrated the presence of lectins binding to glucose and N-acetylglucosamine on the surface of Conidiobolus obscurus spores by using glycosylated serum albumins substituted with fluorescent dyes and colloidal gold. The role of these exocellular lectins was examined in relation to the adhesion of the fungal spores to their host insect and the pathogenicity of the fungus.

Structure and function of the yeast tRNA ligase gene.

We report here the DNA sequence of the entire coding region of the Saccharomyces cerevisiae tRNA ligase gene. tRNA ligase is one of two enzymes required for tRNA splicing in yeast, and the enzyme is likely a single polypeptide with multiple activities. We find that tRNA ligase is a basic protein of 827 amino acids corresponding to a molecular weight of approximately 95,400. The inferred amino acid sequence for tRNA ligase is not significantly homologous to that of other known proteins of similar activity. In addition to the tRNA ligase reading frame and several other unidentified open reading frames, we have found two open reading frames, ORF1 and ORF2, near the 5'-end of the ligase structural gene. One of these, ORF2, produces a divergent transcript which initiates only 125 nucleotides upstream of the tRNA ligase transcript, and is present in approximately the same relative abundance as the transcript for tRNA ligase.