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  • 1
    Electronic Resource
    Electronic Resource
    Dialysable leukocyte extracts modify the course of experimental paracoccidioidomycosis in the Syrian hamster (1993)
    Springer
    Mycopathologia 121 (1993), S. 149-156 
    Details
    ISSN: 1573-0832
    Keywords: Paracoccidioidomycosis ; Syrian hamster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The effect of dialysable leukocyte extracts (DLE) obtained from hamsters immunized withParacoccidioides brasiliensis (immune DLE) and from non-immunized hamsters (non-immune DLE) was studied in hamsters inoculated withP. brasiliensis by the intratesticular route. Treatment with immune or non-immune DLE was started during the third week of infection and was repeated at 7, 11, 15 and 19 weeks. A group of untreated infected animals was used as control. Animals were submitted to the delayed hypersensitivity skin test toP. brasiliensis antigen (PbAg) in vivo and assayed in vitro by the macrophage migration inhibition test in the presence of Phytohemagglutinin (PHA) and PbAg and by immunodiffusion for specific antibody. The animals were sacrificed at 4, 8, 12, 16 and 20 weeks. The morphology and extension of the lesions were studied at the inoculation site, and in lymph nodes, lungs, liver, spleen and kidneys. In contrast to the controls, animals treated with both DLEs maintained a positive cell-mediated immune response throughout the experiment and developed less extensive infection with a significantly lower number of fungi in the lesions. The results suggest that immune and non-immune DLE preparations modified the evolution of experimental paracoccidioidomycosis with equal efficiency. This similarity may be explained by the immunoregulatory activities of both extracts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01104070
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  • 2
    Electronic Resource
    Electronic Resource
    Disruption of the nuclear gene encoding the 20.8-kDa subunit of NADH:ubiquinone reductase ofNeurospora mitochondria (1996)
    Springer
    Molecular genetics and genomics 252 (1996), S. 177-183 
    Details
    ISSN: 1617-4623
    Keywords: Neurospora crassa ; Mitochondria ; Complex I ; Assembly ; Gene disruption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nuclear gene coding for the 20.8-kDa subunit of the membrane arm of respiratory chain NADH:ubiquinone reductase (Complex I) fromNeurospora crassa, nuo-20.8, was localized on linkage group I of the fungal genome. A genomic DNA fragment containing this gene was cloned and a duplication was created in a strain ofN. crassa by transformation. To generate RIP (repeat-induced point) mutations in the duplicated sequence, the transformant was crossed with another strain carrying an auxotrophic marker on chromosome I. To increase the chance of finding an isolate with a non-functionalnuo-20.8 gene, random progeny from the cross were selected against this auxotrophy since RIP of the target gene will only occur in the nucleus carrying the duplication. Among these, we isolated and characterised a mutant strain that lacks the 20.8 kDa mitochondrial protein, indicating that this cysteine-rich polypeptide is not essential. Nevertheless, the absence of the 20.8-kDa subunit prevents the full assembly of complex I. It appears that the peripheral arm and two intermediates of the membrane arm of the enzyme are still formed in the mutant mitochondria. The NADH:ubiquinone reductase activity of sonicated mitochondria from the mutant is rotenone insensitive. Electron microscopy of mutant mitochondria does not reveal any alteration in the structure or numbers of the organelles.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02173218
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  • 3
    Electronic Resource
    Electronic Resource
    Inactivation of the gene coding for the 30.4-kDa subunit of respiratory chain NADH dehydrogenase: is the enzyme essential for Neurospora? (1998)
    Springer
    Molecular genetics and genomics 257 (1998), S. 368-375 
    Details
    ISSN: 1617-4623
    Keywords: Key words Mitochondria ; Complex I ; Gene disruption ; Mutants ; Neurospora crassa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated and characterised the nuclear gene that codes for the 30.4-kDa subunit of the peripheral arm of complex I from Neurospora crassa. The single-copy gene was localised on chromosome VI of the fungal genome by restriction fragment length polymorphism mapping. An extra copy of the gene was introduced into a strain of N. crassa by transformation. This strain was crossed with another strain in order to inactivate, by repeat-induced point mutations, both copies of the duplication carried by the parental transformant. Ascospore progeny from the cross were analysed and a mutant strain lacking the 30.4-kDa protein, nuo30.4, was isolated and further characterised. The mutant appears to assemble the membrane arm of complex I, while formation of the peripheral arm is prevented. Nevertheless, the mutant grows reasonably well – indicating that this well conserved protein is not essential for vegetative growth – and is able to mate with other strains both as male or female. Strains with multiple mutations are readily obtained from heterozygous crosses between different complex I mutants of N. crassa. On the other hand, homozygous crosses between several mutants, including nuo30.4, fail to produce ascospores. These results suggest that complex I plays an essential role during the sexual phase of the life cycle of the fungus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004380050659
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  • 4
    Electronic Resource
    Electronic Resource
    Experimental paracoccidioidomycosis in the Syrian hamster (1982)
    Springer
    Mycopathologia 79 (1982), S. 7-17 
    Details
    ISSN: 1573-0832
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Male hamsters (105) received intratesticular injection of suspension of a live yeast phase culture ofParacoccidioides brasiliensis and were sacrificed weekly during 20 weeks. Humoral immunity was studied by the agar-gel immunodiffusion (ID) and indirect immunofluorescence (IF) tests. Cell-mediated immunity was determined by the macrophage migration inhibition test in the presence of phytohemagglutinin (PHA) andParacoccidioides brasiliensis soluble antigen (PbAg). The morphology of the lesions was studied in the inoculation site, lymph nodes, lung, liver, spleen and kidneys. Disseminated paracoccidioidomycosis was observed in 100% of the animals after the first week. The lesions were initially made up of fungi surrounded by polymorphonuclear neutrophils and macrophages. Up to the 10th week the majority of the lesions appeared as compact confluent ephitelioid granulomas containing rare large fungi, some showing signs of degeneration. At this time, the specific antibody titers and the cellular immune response to PHA and PbAg were highest. From the 11th week on the granulomas became less compact, edematous with the epithelioid cells loosely arranged. This change was accompanied by an increase in the number of fungi showing reproductive activity and was associated with renal amyloidosis and progressive decline of cellular immune response both to PHA and PbAg. Contrariwise the titers of circulating antibodies were maintained. In the present model, disseminated paracoccidioidomycosis of the hamster was associated with depression of cellular immunity, change in the pattern of the granuloma, intense fungi proliferation and amyloidosis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00636175
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  • 5
    Electronic Resource
    Electronic Resource
    The effect of Ketoconazole on experimental paracoccidioidomycosis in the Syrian hamster: immunological and histopathological study (1984)
    Springer
    Mycopathologia 88 (1984), S. 141-148 
    Details
    ISSN: 1573-0832
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of Ketoconazole (KTZ) on the hamster experimental intratesticular paracoccidioidomycosis was studied employing different treatment schedules. KTZ long course treatment beginning at an early stage of the infection was effective in preventing fungal proliferation, dissemination to lymph nodes, spleen and kidneys, and in maintaining low levels of humoral and cellular specific immune responses. KTZ short course treatment starting at an advanced stage of disease resulted in a more severe histopathological picture without significant changes in the immunological profile. The drug prolonged the life span of hamsters infected with Paracoccidioides brasiliensis, but did not prevent mortality. Toxic necrosis of the bone marrow occurred in normal animals receiving 120 mg/kg/day of KTZ but with lower doses no morphologic alterations were observed in heart, lungs, kidneys, adrenals, spleen, liver, intestine or bone marrow.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00436445
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  • 6
    Electronic Resource
    Electronic Resource
    Effect of levamisole on experimental paracoccidioidomycosis in the Syrian hamster: Immunologic and histopathologic correlation (1984)
    Springer
    Mycopathologia 84 (1984), S. 171-180 
    Details
    ISSN: 1573-0832
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The effect of levamisole (LMS) was studied in hamsters inoculated with live yeast phase culture of Paracoccidioides brasiliensis by intratesticular route. One group started LMS therapy at an early stage of infection (LMS3 group), when the animals were immunocompetent, and another group was treated in a later stage, when the immune response was already depressed (LMS12 group). As control, one group was not treated. The alterations induced by levamisole were studied by immunologic and histopathologic parameters. Compared to controls, the LMS3 group presented normal levels of cellular immune response and inflammatory reaction characterized by compact epithelioid granuloma during a longer period of time. In addition, this group showed a lower incidence of amyloidosis and lower fungi proliferation in the lesions. In the LMS12 group a transient enhancement was noteworthy of cellular immune response with maintenance of the compact pattern of the epithelioid granuloma as in the LMS3 group; however, the number of fungi and incidence of amyloidosis were similar to controls. The differences between both treated groups may be accounted for by some factors such as host immune competence, timing and total dose of LMS administered. Levamisole may be of value as additional therapy in paracoccidioidomycosis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00436529
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