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1.
Rev Francoph Lab ; 2022(538): 1, 2022 Jan.
Artigo em Francês | MEDLINE | ID: mdl-34976116
2.
Am J Trop Med Hyg ; 94(3): 611-4, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26787142

RESUMO

We report the case of a French traveler who developed acute pulmonary schistosomiasis 2 months after visiting Benin. He presented with a 1-month history of fever, cough, and thoracic pain. Initial investigations revealed hypereosinophilia and multiple nodular lesions on chest computed tomography scan. Lung biopsies were performed 2 months later because of migrating chest infiltrates and increasing eosinophilia. Histological examination showed schistosomal egg-induced pulmonary granulomas with ova exhibiting a prominent terminal spine, resembling Schistosoma haematobium. However, egg shells were Ziehl-Neelsen positive, raising the possibility of a Schistosoma intercalatum or a Schistosoma guineensis infection. Moreover, involvement of highly infectious hybrid species cannot be excluded considering the atypical early pulmonary oviposition. This case is remarkable because of the rarity of pulmonary schistosomiasis, its peculiar clinical presentation and difficulties in making species identification. It also emphasizes the need to consider schistosomiasis diagnosis in all potentially exposed travelers with compatible symptoms.


Assuntos
Granuloma/parasitologia , Pneumopatias Parasitárias/diagnóstico , Pneumopatias Parasitárias/parasitologia , Schistosoma/isolamento & purificação , Esquistossomose/diagnóstico , Animais , Benin , França , Granuloma/tratamento farmacológico , Humanos , Pneumopatias Parasitárias/tratamento farmacológico , Masculino , Óvulo , Praziquantel/administração & dosagem , Praziquantel/uso terapêutico , Esquistossomose/tratamento farmacológico , Esquistossomose/patologia , Esquistossomicidas/administração & dosagem , Esquistossomicidas/uso terapêutico , Viagem , Adulto Jovem
3.
Am J Trop Med Hyg ; 85(5): 897-9, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22049045

RESUMO

Buruli ulcer is a tropical skin disease caused by Mycobacterium ulcerans. Its mode of transmission is not yet clearly understood. We report here a cutaneous ulcer in a European traveler in South America resulting from a coinfection detected specifically for Mycobacterium ulcerans and Leishmania braziliensis DNA with real-time polymerase chain reaction. This observation of a unique cutaneous ulcer raises the issue about possible modes of transmission of those two pathogens by the same vector.


Assuntos
Úlcera de Buruli/complicações , Coinfecção , Leishmania braziliensis/isolamento & purificação , Leishmaniose Cutânea/complicações , Mycobacterium ulcerans/isolamento & purificação , Adulto , Antibacterianos/uso terapêutico , Antiprotozoários/uso terapêutico , Úlcera de Buruli/tratamento farmacológico , Úlcera de Buruli/epidemiologia , França , Humanos , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/epidemiologia , Masculino , América do Sul/epidemiologia , Viagem
5.
Mycopathologia ; 171(1): 11-21, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20617462

RESUMO

Resistance to 5-fluorocytosine (5-FC) has been poorly investigated in the yeast Candida glabrata. This study was conducted on laboratory mutants obtained by exposure of a wild-type isolate to 5-FC. Based on their susceptibility to 5-fluorouracil (5-FU), two of these mutants were selected for further analysis of the molecular mechanisms of 5-FC resistance. One mutant, resistant to both compounds, exhibited a missense mutation in the gene coding the cytosine deaminase and a decrease in the expression level of the gene coding the uridine monophosphate pyrophosphorylase. The other mutant that showed a reduced susceptibility to 5-FC and 5-FU exhibited an overexpression of the genes coding the thymidylate synthase and a cytosine permease, associated with a missense mutation in the last gene. Thus, beside mutations in the FUR1 gene which represent the most common cause of resistance to 5-FC, other mechanisms may also occur in C. glabrata.


Assuntos
Antifúngicos/farmacologia , Candida glabrata/efeitos dos fármacos , Farmacorresistência Fúngica , Flucitosina/farmacologia , Substituição de Aminoácidos , Citosina Desaminase/genética , Análise Mutacional de DNA , Fluoruracila/farmacologia , Expressão Gênica , Mutação de Sentido Incorreto , Pentosiltransferases/biossíntese , Timidilato Sintase/biossíntese
6.
Mycopathologia ; 170(3): 161-4, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20340045

RESUMO

During the past two decades, an increasing number of unusual moulds has been reported as responsible for septicaemia and systemic or disseminated infections in immunocompromised patients. Investigation of fever in a 10-year-old boy with acute myeloblastic leukaemia, including blood cultures on selective media, allowed the diagnosis of a fungaemia due to the slow-growing fungus Acremonium strictum. The patient recovered with liposomal amphotericin B (AmB) and voriconazole, followed by voriconazole alone due to AmB resistance. Facing a neutropenic patient with fever, clinicians usually suspect bacterial or viral aetiologies. This case, however, illustrates the need for mycological analysis of blood samples in febrile neutropenic patients and for antifungal susceptibility testing.


Assuntos
Acremonium/isolamento & purificação , Fungemia/diagnóstico , Leucemia Mieloide Aguda/complicações , Anfotericina B/uso terapêutico , Antifúngicos/uso terapêutico , Sangue/microbiologia , Criança , Febre/etiologia , Fungemia/tratamento farmacológico , Fungemia/microbiologia , Humanos , Hospedeiro Imunocomprometido , Leucemia Mieloide Aguda/tratamento farmacológico , Masculino , Neutropenia/etiologia , Pirimidinas/uso terapêutico , Triazóis/uso terapêutico , Voriconazol
7.
BMC Microbiol ; 9: 177, 2009 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-19703288

RESUMO

BACKGROUND: Aspergillus fumigatus is the most common agent of invasive aspergillosis, a feared complication in severely immunocompromised patients. Despite the recent commercialisation of new antifungal drugs, the prognosis for this infection remains uncertain. Thus, there is a real need to discover new targets for therapy. Particular attention has been paid to the biochemical composition and organisation of the fungal cell wall, because it mediates the host-fungus interplay. Conidia, which are responsible for infections, have melanin as one of the cell wall components. Melanin has been established as an important virulence factor, protecting the fungus against the host's immune defences. We suggested that it might also have an indirect role in virulence, because it is required for correct assembly of the cell wall layers of the conidia. RESULTS: We used three A. fumigatus isolates which grew as white or brown powdery colonies, to demonstrate the role of melanin. Firstly, sequencing the genes responsible for biosynthesis of melanin (ALB1, AYG1, ARP1, ARP2, ABR1 and ABR2) showed point mutations (missense mutation, deletion or insertion) in the ALB1 gene for pigmentless isolates or in ARP2 for the brownish isolate. The isolates were then shown by scanning electron microscopy to produce numerous, typical conidial heads, except that the conidia were smooth-walled, as previously observed for laboratory mutants with mutations in the PKSP/ALB1 gene. Flow cytometry showed an increase in the fibronectin binding capacity of conidia from mutant isolates, together with a marked decrease in the binding of laminin to the conidial surface. A marked decrease in the electronegative charge of the conidia and cell surface hydrophobicity was also seen by microelectrophoresis and two-phase partitioning, respectively. Ultrastructural studies of mutant isolates detected considerable changes in the organisation of the conidial wall, with the loss of the outermost electron dense layer responsible for the ornamentations seen on the conidial surface in wild-type strains. Finally, analysis of the conidial surface of mutant isolates by atomic force microscopy demonstrated the absence of the outer cell wall rodlet layer which is composed of hydrophobins. CONCLUSION: These results suggest that, in addition to a protective role against the host's immune defences, melanin is also a structural component of the conidial wall that is required for correct assembly of the cell wall layers and the expression at the conidial surface of adhesins and other virulence factors.


Assuntos
Aspergillus fumigatus/genética , Parede Celular/química , Melaninas/biossíntese , Esporos Fúngicos/ultraestrutura , Aspergilose/microbiologia , Aspergillus fumigatus/patogenicidade , Aspergillus fumigatus/ultraestrutura , Parede Celular/ultraestrutura , DNA Fúngico/genética , Citometria de Fluxo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Microscopia Eletrônica de Varredura , Análise de Sequência de DNA , Esporos Fúngicos/genética , Virulência/genética
8.
Antimicrob Agents Chemother ; 53(7): 3034-41, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19380598

RESUMO

Petite mutations have been described in Saccharomyces cerevisiae and pathogenic yeasts. However, previous studies of the phenotypic traits of these petite mutants reported that they express azole resistance. We describe a clinical isolate of Candida glabrata with a striking association between increased susceptibility to azoles and respiratory deficiency. This isolate was obtained from a urine sample together with a respiration-competent C. glabrata isolate which exhibited azole resistance. The respiratory status of the two isolates was confirmed by cultivation on glycerol-containing agar and oxygraphy. Flow cytometry revealed the normal incorporation of rhodamine 123, and mitochondrial sections with typical cristae were seen by transmission electron microscopy for both isolates. Together, these results suggested a nuclear origin for the reduced respiratory capacity of the hypersusceptible isolate. The sterol contents of these isolates were similar to the sterol content of a reference strain. Sequencing of the ERG11 and PDR1 genes revealed that the sequences were identical in the two isolates, demonstrating their close relatedness. In addition to silent mutations, they carried a nonsense mutation in PDR1 that led to the truncation of transcription factor Pdr1p. They also overexpressed both PDR1 and one of its targets, CDR1, providing a possible explanation for the azole resistance of the respiration-competent isolate. In conclusion, in addition to azole resistance, which is a common feature of C. glabrata mitochondrial petite mutants, the mutation of a nuclear gene affecting aerobic growth may lead to azole hypersusceptibility; however, the mechanisms underlying this phenotype remain to be determined.


Assuntos
Antifúngicos/farmacologia , Azóis/farmacologia , Candida glabrata/efeitos dos fármacos , Candida glabrata/crescimento & desenvolvimento , Adulto , Aerobiose , Candida glabrata/metabolismo , Candida glabrata/ultraestrutura , Cromatografia Líquida de Alta Pressão , Ergosterol/metabolismo , Feminino , Citometria de Fluxo , Genes Fúngicos/genética , Genes Fúngicos/fisiologia , Humanos , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Transmissão , Dados de Sequência Molecular , Polienos/farmacologia
9.
Med Mycol ; 47(4): 387-97, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19107638

RESUMO

The colonization of airways by filamentous fungi and the development of respiratory infections require some predisposing factors as encountered in patients with cystic fibrosis (CF). Indeed, the defective mucociliary clearance which characterizes the disease is associated with local immunological disorders. In addition, the prolonged therapy with antibiotics and the use of corticosteroid treatments also facilitate fungal growth. An important fungal biota has been described in respiratory secretions of patients suffering from CF. Aspergillus fumigatus, Scedosporium apiospermum and Aspergillus terreus for filamentous fungi and Candida albicans for yeasts are the main fungal species associated with CF. Although less common, several fungal species including Aspergillus flavus and Aspergillus nidulans may be isolated transiently from CF respiratory secretions, while others such as Exophiala dermatitidis and Scedosporium prolificans may chronically colonize the airways. Moreover, some of them like Penicillium emersonii and Acrophialophora fusispora are encountered in humans almost exclusively in the context of CF. As fungal complications in CF patients are essentially caused by filamentous fungi the present review will not include works related to yeasts. In CF patients, fungi may sometimes be responsible for deterioration of lung function, as occurs in allergic broncho-pulmonary aspergillosis (ABPA) which is the most common fungal disease in this context. Additionally, although the clinical relevance of the fungal airway colonization is still a matter of debate, filamentous fungi may contribute to the local inflammatory response, and therefore to the progressive deterioration of the lung function.


Assuntos
Fibrose Cística/complicações , Fungos/classificação , Fungos/isolamento & purificação , Pneumopatias Fúngicas/epidemiologia , Pneumopatias Fúngicas/microbiologia , Humanos , Prevalência
10.
Antimicrob Agents Chemother ; 52(10): 3701-9, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18694952

RESUMO

Unlike the molecular mechanisms that lead to azole drug resistance, the molecular mechanisms that lead to polyene resistance are poorly documented, especially in pathogenic yeasts. We investigated the molecular mechanisms responsible for the reduced susceptibility to polyenes of a clinical isolate of Candida glabrata. Sterol content was analyzed by gas-phase chromatography, and we determined the sequences and levels of expression of several genes involved in ergosterol biosynthesis. We also investigated the effects of the mutation harbored by this isolate on the morphology and ultrastructure of the cell, cell viability, and vitality and susceptibility to cell wall-perturbing agents. The isolate had a lower ergosterol content in its membranes than the wild type, and the lower ergosterol content was found to be associated with a nonsense mutation in the ERG6 gene and induction of the ergosterol biosynthesis pathway. Modifications of the cell wall were also seen, accompanied by increased susceptibility to cell wall-perturbing agents. Finally, this mutation, which resulted in a marked fitness cost, was associated with a higher rate of cell mortality. Wild-type properties were restored by complementation of the isolate with a centromeric plasmid containing a wild-type copy of the ERG6 gene. In conclusion, we have identified the molecular event responsible for decreased susceptibility to polyenes in a clinical isolate of C. glabrata. The nonsense mutation detected in the ERG6 gene of this isolate led to a decrease in ergosterol content. This isolate may constitute a useful tool for analysis of the relevance of protein trafficking in the phenomena of azole resistance and pseudohyphal growth.


Assuntos
Antifúngicos/farmacologia , Candida glabrata/efeitos dos fármacos , Candida glabrata/genética , Códon sem Sentido , Genes Fúngicos , Polienos/farmacologia , Azóis/farmacologia , Sequência de Bases , Candida glabrata/isolamento & purificação , Candida glabrata/metabolismo , Candidíase/tratamento farmacológico , Candidíase/microbiologia , DNA Fúngico/genética , Farmacorresistência Fúngica/genética , Ergosterol/metabolismo , Guanina/análogos & derivados , Humanos , Dados de Sequência Molecular
11.
Trans R Soc Trop Med Hyg ; 102(4): 389-93, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18295290

RESUMO

Tinea imbricata, also known as 'Tokelau', is an uncommon superficial mycosis caused by the anthropophilic dermatophyte Trichophyton concentricum. Cutaneous lesions appear characteristically as scaly and concentric rings that may cover all parts of the body. Often acquired in childhood, tinea imbricata is a chronic disease and lichenification is extremely common due to pruritus. The dermatophytosis mainly occurs in the South Pacific, but also in some regions of Southeast Asia and Central or South America. Tinea imbricata usually affects people living in primitive and isolated conditions. Mycological analysis is required for the diagnosis. The epidemiological and mycological study reported here took place in the Solomon Islands from June-September 2006. Skin scrapings were collected from 29 Melanesian patients (aged 8 months to 58 years) with chronic cutaneous lesions and were analysed mycologically in the Laboratory of Parasitology and Mycology of Angers University Hospital (France). Ten patients showed very evocative lesions with a positive direct examination, but T. concentricum was only isolated from three patients. Identification of the strains was confirmed by sequencing of the internal transcribed spacer (ITS) regions. With the increase in international travel, one cannot disregard that this very rare species may be isolated by mycologists in temperate areas from patients coming from endemic foci.


Assuntos
Tinha/microbiologia , Trichophyton/isolamento & purificação , Adolescente , Adulto , Criança , Pré-Escolar , Doença Crônica , Feminino , Humanos , Lactente , Masculino , Melanesia/epidemiologia , Pessoa de Meia-Idade , Técnicas de Tipagem Micológica/métodos , Tinha/epidemiologia , Tinha/patologia , Trichophyton/classificação
12.
Microbes Infect ; 9(5): 558-65, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17395518

RESUMO

A Cu,Zn-superoxide dismutase has been characterized from Scedosporium apiospermum, a fungus which often colonizes the respiratory tract of patients with cystic fibrosis. Enzyme production was stimulated by iron starvation. Purification was achieved from mycelial extract from 7-day-old cultures on Amberlite XAD-16. The purified enzyme presented a relative molecular mass of 16.4 kDa under reducing conditions and was inhibited by potassium cyanide and diethyldithiocarbamate, which are two known inhibitors of Cu,Zn-SODs. Its optimum pH was 7.0 and the enzyme retained full activity after pretreatment at temperatures up to 50 degrees C. Moreover, a 450-bp fragment of the gene encoding the enzyme was amplified by PCR using degenerate primers designed from sequence alignment of four fungal Cu,Zn-SODs. Sequence data from this fragment allowed us to design primers which were used to amplify by walking-PCR the flanking regions of the known fragment. SaSODC gene (890 bp) corresponded to a 154 amino acid polypeptide with a predicted molecular mass of 15.9 kDa. A database search for sequence homology revealed for the deduced amino acid sequence 72 and 83% identity rate with Cu,Zn-SODs from Aspergillus fumigatus and Neurospora crassa, respectively. To our knowledge, this enzyme is the first putative virulence factor of S. apiospermum to be characterized.


Assuntos
Scedosporium/enzimologia , Scedosporium/genética , Superóxido Dismutase/genética , Sequência de Aminoácidos , Clonagem Molecular , Cobre/metabolismo , Análise de Sequência de DNA , Superóxido Dismutase/metabolismo , Zinco/metabolismo
13.
Antimicrob Agents Chemother ; 51(3): 982-90, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17158937

RESUMO

Little information is available about the molecular mechanisms responsible for polyene resistance in pathogenic yeasts. A clinical isolate of Candida glabrata with a poor susceptibility to polyenes, as determined by disk diffusion method and confirmed by determination of MIC, was recovered from a patient treated with amphotericin B. Quantitative analysis of sterols revealed a lack of ergosterol and an accumulation of late sterol intermediates, suggesting a defect in the final steps of the ergosterol pathway. Sequencing of CgERG11, CgERG6, CgERG5, and CgERG4 genes revealed exclusively a unique missense mutation in CgERG6 leading to the substitution of a cysteine by a phenylalanine in the corresponding protein. In addition, real-time reverse transcription-PCR demonstrated an overexpression of genes encoding enzymes involved in late steps of the ergosterol pathway. Moreover, this isolate exhibited a pseudohyphal growth whatever the culture medium used, and ultrastructural changes of the cell wall of blastoconidia were seen consisting in a thinner inner layer. Cell wall alterations were also suggested by the higher susceptibility of growing cells to Calcofluor white. Additionally, complementation of this isolate with a wild-type copy of the CgERG6 gene restored susceptibility to polyenes and a classical morphology. Together, these results demonstrated that mutation in the CgERG6 gene may lead to a reduced susceptibility to polyenes and to a pseudohyphal growth due to the subsequent changes in sterol content of the plasma membrane.


Assuntos
Antifúngicos/farmacologia , Candida glabrata/efeitos dos fármacos , Candidíase/microbiologia , Metiltransferases/genética , Mutação de Sentido Incorreto/fisiologia , Polienos/farmacologia , Candida glabrata/genética , Candida glabrata/crescimento & desenvolvimento , Primers do DNA , Genes Fúngicos/genética , Teste de Complementação Genética , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Fenótipo , RNA Fúngico/biossíntese , RNA Fúngico/genética , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Esteróis/metabolismo
14.
Antimicrob Agents Chemother ; 49(11): 4608-15, 2005 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16251302

RESUMO

Azole resistance has been insufficiently investigated in the yeast Candida tropicalis. Here we determined the molecular mechanisms responsible for azole resistance in a clinical isolate of this pathogenic yeast. Antifungal susceptibility testing performed by a disk diffusion method showed resistance or markedly decreased susceptibility to azoles, which was confirmed by determination of MICs. Considering the relationship between azole susceptibility and the respiration reported for other yeast species, the respiratory activity of this isolate was investigated. Flow cytometry using rhodamine 123 and oxygraphy demonstrated an increased respiratory activity, which was not linked to an overexpression or increased number of copies of the mitochondrial genome. Among previously described resistance mechanisms, an increased activity of efflux pumps was investigated by flow cytometry using rhodamine 6G. However, the efflux of rhodamine 6G was lower in the resistant isolate than in susceptible ones. Likewise, real-time reverse transcription-PCR quantification of the expression of C. tropicalis MDR1 (CtMDR1), which encodes an efflux protein belonging to the major facilitator superfamily, did not show overexpression of this gene. In contrast, the resistant isolate overexpressed the CtERG11 gene coding for lanosterol 14alpha-demethylase. This was in agreement with the larger amount of ergosterol found in this isolate. Moreover, sequencing of CtERG11 showed a point mutation leading to a tyrosine substitution in the protein sequence, which might lead to decreased binding affinity for azoles. In conclusion, overexpression of CtERG11 associated with a missense mutation in this gene seemed to be responsible for the acquired azole resistance of this clinical isolate.


Assuntos
Azóis/farmacologia , Candida/efeitos dos fármacos , Farmacorresistência Fúngica/genética , Sequência de Bases , Candida/genética , Candida/metabolismo , Sistema Enzimático do Citocromo P-450/genética , Genes MDR , Humanos , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Mutação de Sentido Incorreto , Oxirredutases/genética , Consumo de Oxigênio/efeitos dos fármacos , Rodaminas/metabolismo , Esterol 14-Desmetilase , Esteróis/análise
15.
Sante ; 15(3): 195-9, 2005.
Artigo em Francês | MEDLINE | ID: mdl-16207583

RESUMO

We report a case of disseminated African histoplasmosis with lymph node and digestive involvement in a 19-year-old man living in the Kayes district of Mali. The patient, HIV-seronegative and not otherwise immunocompromised, presented voluminous cervical and axillary adenopathies as well as retrosternal and mesenteric tumor lesions. Direct examination of biopsy tissue showed numerous specimens of Histoplasma capsulatum var. duboisii. Because direct fungal techniques are the easiest and the most effective method of diagnostic investigation, no cultures were performed. Intolerance to therapy with amphotericin b and ketoconazole led its rapid replacement by surgical treatment: partial excision of the abdominal lesions led to partial remission of the symptoms.


Assuntos
Histoplasmose/diagnóstico , Adulto , Histoplasma/classificação , Humanos , Doenças Linfáticas/microbiologia , Masculino , Mali , Peritonite/microbiologia , Abscesso Subfrênico/microbiologia
16.
Mycopathologia ; 160(2): 117-23, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16170606

RESUMO

Here we report a case of cutaneous alternariosis in a 74-year-old man treated by corticotherapy for myasthenia, and presenting with papular, crusted lesions on the left elbow and the right knee. Histological examination of the biopsy specimens showed fungal hyphae associated with round-shaped cells which were highly suggestive of alternariosis. Mycological culture allowed the isolation of a dematiaceous fungus which was identified as a member of the Alternaria infectoria species-group. This was confirmed by PCR amplification and sequencing of the internal transcribed spacer domain of the gene encoding nuclear ribosomal DNA and of the mitochondrial small subunit ribosomal DNA domain. The fungus was therefore referred to the Scientific Institute of Public Health where it was identified as Alternaria infectoria, on the basis of its very small 1 or 2-celled conidia often arranged in long chains and presenting with very long secondary conidiophores. Corticotherapy was stopped and a local antifungal treatment with ketoconazole was initiated, allowing the stabilisation of the cutaneous lesions within 2 months.


Assuntos
Alternaria/classificação , Alternaria/isolamento & purificação , Dermatomicoses/microbiologia , Adulto , Idoso , Alternaria/genética , Alternaria/ultraestrutura , DNA Mitocondrial/genética , DNA Espaçador Ribossômico/análise , Dermatomicoses/diagnóstico , Dermatomicoses/patologia , Feminino , Humanos , Joelho/patologia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA
17.
J Antimicrob Chemother ; 56(2): 307-14, 2005 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-15958498

RESUMO

OBJECTIVES: To define the pathogenicity of respiration-deficient mutants of Candida glabrata, which present a reduced susceptibility to azoles, that are easily induced in vitro by exposure to these drugs or to ethidium bromide and that may be selected in vivo in patients receiving fluconazole. METHODS: Two wild-type isolates of C. glabrata were compared with their respective fluconazole- or ethidium bromide-induced petite mutants, regarding the carbohydrate and protein composition of the cell wall, as well as their surface physical properties, and also their adherence abilities and virulence in mice. RESULTS: Flow cytometric analysis of cell wall carbohydrates using several fluorescent lectins showed an increased binding of mutant cells to concanavalin A compared with their parent isolates, suggesting a greater availability or an increased amount of glucose-mannose residues at the cell surface in petite mutants. Likewise, some quantitative differences between parent and mutant isolates were shown by SDS-PAGE in protein extracts from blastoconidia. Regarding the surface physical properties, no significant differences were seen in the electrophoretic mobility determined by microelectrophoresis, but the two-phase partitioning method revealed a lower cell surface hydrophobicity for petite mutants. Moreover, mutant cells exhibited significant overexpression of CgEPA1 as revealed by real-time reverse transcription-PCR, but the adherence capacities to Caco-2 cells, a human enterocyte line, were not significantly different. Finally, in agreement with their slower growth, petite mutants were less virulent than parent isolates in a murine model of systemic infection. CONCLUSION: This low virulence in mice suggests that petite mutants could be disregarded clinically although they may arise during fluconazole therapy.


Assuntos
Aderência Bacteriana/genética , Candida glabrata/genética , Candidíase/microbiologia , Parede Celular/metabolismo , Proteínas Fúngicas/genética , Lectinas/genética , Animais , Encéfalo/microbiologia , Células CACO-2 , Candida glabrata/patogenicidade , Contagem de Colônia Microbiana , Modelos Animais de Doenças , Eletroforese em Gel de Poliacrilamida , Feminino , Citometria de Fluxo , Proteínas Fúngicas/metabolismo , Humanos , Rim/microbiologia , Lectinas/metabolismo , Fígado/microbiologia , Camundongos , Mutação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Virulência
18.
J Clin Microbiol ; 43(3): 1484-7, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15750142

RESUMO

Acrophialophora fusispora is a thermotolerant soil fungus which is very unusual in clinical samples. Here we report four cases of transient or chronic airway colonization by A. fusispora in patients with cystic fibrosis (CF). However, the prevalence of this fungus in CF patients may be underestimated due to the currently poor knowledge of this fungus in most of the medical mycology laboratories. In addition, its clinical importance regarding CF remains to be evaluated.


Assuntos
Brônquios/microbiologia , Fibrose Cística/microbiologia , Fungos Mitospóricos/isolamento & purificação , Microbiologia do Solo , Escarro/microbiologia , Adolescente , Adulto , Pré-Escolar , Feminino , Humanos , Masculino
19.
Microb Pathog ; 37(3): 131-40, 2004 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-15351036

RESUMO

The adherence of Sporothrix schenckii yeast cells to several extracellular matrix (ECM) components has already been demonstrated, but the mechanisms of these interactions remained to be defined. In indirect immunofluorescence assays with polyclonal antibodies directed towards the ECM proteins, both hyphae and yeast cells of S. schenckii exhibited the ability to bind laminin and fibronectin. Flow cytometry confirmed the binding of these proteins, and revealed a significant greater binding capability for the yeast cells than for the conidia. Fibronectin and laminin binding was dose-dependent and specific. In addition, competition experiments with synthetic peptides mimicking the adhesive sequences of these proteins, or with cell wall fractions and carbohydrates constitutive of their sugar chains, were performed in order to specify the peptide or carbohydrate motifs involved in the recognition process. A 50% reduction was noticed in fibronectin binding in the presence of the synthetic peptide RGD, and a 38% reduction in laminin binding with the peptide YIGSR. Some carbohydrate-containing fractions of the yeast cell wall also inhibited the binding of fibronectin, but had no significant effect on laminin binding. Together, these results suggest the presence at the yeast surface of distinct receptors for laminin and fibronectin.


Assuntos
Fibronectinas/metabolismo , Laminina/metabolismo , Sporothrix/metabolismo , Adesão Celular , Fibronectinas/química , Citometria de Fluxo , Imunofluorescência , Laminina/química , Peptídeos/síntese química , Peptídeos/química , Peptídeos/farmacologia , Ligação Proteica , Sporothrix/crescimento & desenvolvimento , Sporothrix/fisiologia
20.
Antimicrob Agents Chemother ; 48(5): 1788-96, 2004 May.
Artigo em Inglês | MEDLINE | ID: mdl-15105136

RESUMO

We previously showed that resistant colonies of Candida glabrata inside the azole inhibition zones had respiratory deficiency due to mutations in mitochondrial DNA. Here, we analyzed the mechanisms of azole resistance in petite mutants of C. glabrata obtained by exposure to fluconazole or induced by ethidium bromide. The respiratory deficiency of these mutants was confirmed by oxygraphy and flow cytometric analysis with rhodamine 123, and its mitochondrial origin was demonstrated by transmission electron microscopy and restriction endonuclease analysis of the mitochondrial DNA. Flow cytometry with rhodamine 6G suggested an increased drug efflux in mutant cells, which was further supported by Northern blot analysis of the expression of the C. glabrata CDR1 (CgCDR1) and CgCDR2 genes, encoding efflux pumps. Conversely, the expression of CgERG11, which encodes the azole target, was not affected by petite mutations, and no differences were seen in the sequence of this gene between parent isolates and mutants. Moreover, sterol analysis showed similar overall amount of sterols in parent and mutant cells, but quantitative modifications were observed in the mutants, with almost undetectable biosynthesis intermediates. Further analysis performed after separation of free sterols from steryl esters revealed a defect in sterol esterification in mutant cells, with free ergosterol representing 92% of the overall sterol content. Thus, resistance or decreased susceptibility to azoles in petite mutants of C. glabrata is associated with increased expression of CgCDR1 and, to a lesser extent, of CgCDR2. In addition, the marked increase in free ergosterol content would explain their increased susceptibility to polyenes.


Assuntos
Antifúngicos/farmacologia , Azóis/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/genética , Northern Blotting , DNA Mitocondrial/genética , Farmacorresistência Fúngica Múltipla , Citometria de Fluxo , Fluconazol/farmacologia , Corantes Fluorescentes , Proteínas Fúngicas/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Testes de Sensibilidade Microbiana , Microscopia Eletrônica , Mutação/fisiologia , Rodaminas , Esteróis/metabolismo
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