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Proteomics-Based Characterization of the Humoral Immune Response in Sporotrichosis: Toward Discovery of Potential Diagnostic and Vaccine Antigens  [PDF]
Anderson Messias Rodrigues?,Geisa Ferreira Fernandes?,Leticia Mendes Araujo?,Paula Portella Della Terra?,Priscila Oliveira dos Santos?,Sandro Antonio Pereira?,Tania Maria Pacheco Schubach?,Eva Burger?,Leila Maria Lopes-Bezerra?,Zoilo Pires de Camargo
PLOS Neglected Tropical Diseases , 2015, DOI: 10.1371/journal.pntd.0004016
Abstract: Background Sporothrix schenckii and associated species are agents of human and animal sporotrichosis that cause large sapronoses and zoonoses worldwide. Epidemiological surveillance has highlighted an overwhelming occurrence of the highly pathogenic fungus Sporothrix brasiliensis during feline outbreaks, leading to massive transmissions to humans. Early diagnosis of feline sporotrichosis by demonstrating the presence of a surrogate marker of infection can have a key role for selecting appropriate disease control measures and minimizing zoonotic transmission to humans. Methodology We explored the presence and diversity of serum antibodies (IgG) specific against Sporothrix antigens in cats with sporotrichosis and evaluated the utility of these antibodies for serodiagnosis. Antigen profiling included protein extracts from the closest known relatives S. brasiliensis and S. schenckii. Enzyme-linked immunosorbent assays and immunoblotting enabled us to characterize the major antigens of feline sporotrichosis from sera from cats with sporotrichosis (n = 49), healthy cats (n = 19), and cats with other diseases (n = 20). Principal Findings Enzyme-linked immunosorbent assay-based quantitation of anti-Sporothrix IgG exhibited high sensitivity and specificity in cats with sporotrichosis (area under the curve, 1.0; 95% confidence interval, 0.94–1; P<0.0001) versus controls. The two sets of Sporothrix antigens were remarkably cross-reactive, supporting the hypothesis that antigenic epitopes may be conserved among closely related agents. One-dimensional immunoblotting indicated that 3-carboxymuconate cyclase (a 60-kDa protein in S. brasiliensis and a 70-kDa protein in S. schenckii) is the immunodominant antigen in feline sporotrichosis. Two-dimensional immunoblotting revealed six IgG-reactive isoforms of gp60 in the S. brasiliensis proteome, similar to the humoral response found in human sporotrichosis. Conclusions A convergent IgG-response in various hosts (mice, cats, and humans) has important implications for our understanding of the coevolution of Sporothrix and its warm-blooded hosts. We propose that 3-carboxymuconate cyclase has potential for the serological diagnosis of sporotrichosis and as target for the development of an effective multi-species vaccine against sporotrichosis in animals and humans.
Sporotrichosis in Rio de Janeiro, Brazil: Sporothrix brasiliensis Is Associated with Atypical Clinical Presentations  [PDF]
Rodrigo Almeida-Paes ,Manoel Marques Evangelista de Oliveira,Dayvison Francis Saraiva Freitas,Ant?nio Carlos Francesconi do Valle,Rosely Maria Zancopé-Oliveira,Maria Clara Gutierrez-Galhardo
PLOS Neglected Tropical Diseases , 2014, DOI: 10.1371/journal.pntd.0003094
Abstract: Background There have been several recent changes in the taxonomy of Sporothrix schenckii as well as new observations regarding the clinical aspects of sporotrichosis. In this study, we determined the identification of the Sporothrix species associated with both classic and unusual clinical aspects of sporotrichosis observed in the endemic area of sporotrichosis in Rio de Janeiro, Brazil. Methodology/Principal Findings To verify whether S. brasiliensis is associated with clinical manifestations of sporotrichosis, a cross-sectional study was performed in which Sporothrix isolates from 50 patients with different clinical manifestations were analyzed and their isolates were studied by phenotypic and genotypic methods. Data from these patients revealed a distinct clinical picture and therapeutic response in infections caused by Sporothrix brasiliensis (n = 45) compared to patients with S. schenckii sensu stricto (n = 5). S. brasiliensis was associated with disseminated cutaneous infection without underlying disease, hypersensitivity reactions, and mucosal infection, whereas patients with S. schenckii presented with less severe and more often localized disease, similar to the majority of previously described sporotrichosis cases. Interestingly, S. brasiliensis-infected patients overall required shorter durations of itraconazole (median 16 weeks) compared to the individuals with S. schenckii (median 24 weeks). Conclusions/Significance These findings suggest that Sporothrix species are linked to different clinical manifestations of sporotrichosis and that S. brasiliensis is effectively treated with oral itraconazole.
Ultrastructural study of the mycelial phase of clinical isolates of Sporothrix schenckii obtained from feline, canine and human cases of sporotrichosis
Madrid, Isabel Martins;Mattei, Antonella Souza;Soares, Mauro Pereira;Nobre, Márcia de Oliveira;Meireles, Mário Carlos Araújo;
Brazilian Journal of Microbiology , 2011, DOI: 10.1590/S1517-83822011000300037
Abstract: using transmission electron microscopy, we studied the presence of melanin and cell wall thickness of clinical isolates of sporothrix schenckii obtained from cats, dogs and humans as compared to reference strains. we detected differences regarding presence of the melanin among the clinical isolates of s. schenckii and a correlation between presence of melanin and cell wall thickness.
Sporothrix schenckii and sporotrichosis
Lopes-Bezerra, Leila M.;Schubach, Armando;Costa, Rosane O.;
Anais da Academia Brasileira de Ciências , 2006, DOI: 10.1590/S0001-37652006000200009
Abstract: for a long time sporotrichosis has been regarded to have a low incidence in brazil; however, recent studies demonstrate that not only the number of reported cases but also the incidence of more severe or atypical clinical forms of the disease are increasing. recent data indicate that these more severe forms occur in about 10% of patients with confirmed diagnosis. the less frequent forms, mainly osteoarticular sporotrichosis, might be associated both with patient immunodepression and zoonotic transmission of the disease. the extracutaneous form and the atypical forms are a challenge to a newly developed serological test, introduced as an auxiliary tool for the diagnosis of unusual clinical forms of sporotrichosis.
Sporothrix schenckii and sporotrichosis  [cached]
Lopes-Bezerra Leila M.,Schubach Armando,Costa Rosane O.
Anais da Academia Brasileira de Ciências , 2006,
Abstract: For a long time sporotrichosis has been regarded to have a low incidence in Brazil; however, recent studies demonstrate that not only the number of reported cases but also the incidence of more severe or atypical clinical forms of the disease are increasing. Recent data indicate that these more severe forms occur in about 10% of patients with confirmed diagnosis. The less frequent forms, mainly osteoarticular sporotrichosis, might be associated both with patient immunodepression and zoonotic transmission of the disease. The extracutaneous form and the atypical forms are a challenge to a newly developed serological test, introduced as an auxiliary tool for the diagnosis of unusual clinical forms of sporotrichosis.
ESPOROTRICOSE FELINA: RELATO DE CASOS SCIENTIFIC REPORT:FELINE SPOROTRICHOSIS – CASE REPORTS  [cached]
Ana Raquel Mano Meinerz,Patrícia da Silva Nascente,Luiz Filipe Damé Schuch,Renata Osório de Faria
Ciência Animal Brasileira , 2007,
Abstract: o felino doméstico com esporotricose representa um papel importante na transmiss o do S. schenckii a ou-tros animais e para o homem sendo crescente o aumento do número de casos zoonóticos da micose. o estudo des-creve três casos de esporotricose em gatos semidomicilia-dos, convivendo no mesmo bairro. os felinos apresenta-ram les es cutaneas ulceradas e crostosas, sendo realizada a confirma o do diagnóstico após a análise micológica com o isolamento do S. schenckii. o estudo ressalta a im-portancia dos felinos domésticos como propagadores da esporotricose para animais e para o homem. PALAVRAS-CHAVE: Esporotricose, zoonose, felino, Sporothrix schenckii. the domestic feline with sporotrichosis represents an important role in the transmission of the S. schenckii schenckii to other animal species and for the man being growing the increase of the number of zoonotic cases of the mycosis. the study reports three cases of feline sporo-trichosis in semi-domiciled cats, living together in the same neighborhood. the feline presented cutaneous ulcerated and crusted injuries being accomplished the confirmation of the diagnosis after the analysis micológica with the isolation of S. schenckii. the study emphasizes the importance of the domestic feline as diffuser of the sporotrichosis for other animals and for the man itself. KEY-WORDS: feline, sporotrichosis, zoonosis, Sporothrix schenckii.
Differences in Cell Morphometry, Cell Wall Topography and Gp70 Expression Correlate with the Virulence of Sporothrix brasiliensis Clinical Isolates  [PDF]
Rafaela A. Castro, Paula H. Kubitschek-Barreira, Pedro A. C. Teixeira, Glenda F. Sanches, Marcus M. Teixeira, Leonardo P. Quintella, Sandro R. Almeida, Rosane O. Costa, Zoilo P. Camargo, Maria S. S. Felipe, Wanderley de Souza, Leila M. Lopes-Bezerra
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0075656
Abstract: Sporotrichosis is a chronic infectious disease affecting both humans and animals. For many years, this subcutaneous mycosis had been attributed to a single etiological agent; however, it is now known that this taxon consists of a complex of at least four pathogenic species, including Sporothrix schenckii and Sporothrix brasiliensis. Gp70 was previously shown to be an important antigen and adhesin expressed on the fungal cell surface and may have a key role in immunomodulation and host response. The aim of this work was to study the virulence, morphometry, cell surface topology and gp70 expression of clinical isolates of S. brasiliensis compared with two reference strains of S. schenckii. Several clinical isolates related to severe human cases or associated with the Brazilian zoonotic outbreak of sporotrichosis were genotyped and clustered as S. brasiliensis. Interestingly, in a murine subcutaneous model of sporotrichosis, these isolates showed a higher virulence profile compared with S. schenckii. A single S. brasiliensis isolate from an HIV-positive patient not only showed lower virulence but also presented differences in cell morphometry, cell wall topography and abundant gp70 expression compared with the virulent isolates. In contrast, the highly virulent S. brasiliensis isolates showed reduced levels of cell wall gp70. These observations were confirmed by the topographical location of the gp70 antigen using immunoelectromicroscopy in both species. In addition, the gp70 molecule was sequenced and identified using mass spectrometry, and the sequenced peptides were aligned into predicted proteins using Blastp with the S. schenckii and S. brasiliensis genomes.
Suscetibilidade in vitro de isolados de Sporothrix schenckii frente à terbinafina e itraconazol
Meinerz, Ana Raquel Mano;Nascente, Patrícia da Silva;Schuch, Luiz Filipe Damé;Cleff, Marlete Brum;Santin, Rosema;Brum, Cristiane da Silva;Nobre, Márcia de Oliveira;Meireles, Mario Carlos Araújo;Mello, Jo?o Roberto de Braga;
Revista da Sociedade Brasileira de Medicina Tropical , 2007, DOI: 10.1590/S0037-86822007000100012
Abstract: the study objective was to determine the in vitro activity of terbinafine and itraconazole through the microdilution technique in broth (ncclsm27-a2), adapted for dimorphic fungus, in relation to 12 isolates of sporothrix schenckii. six were from feline sporotrichosis, three from human sporotrichosis, one from a dog and two from human isolates originating from instituto oswaldo cruz. the inoculum and antifungal concentrates were distributed on microplates that were incubated at 35°c for five days. minimum inhibitory concentration readings were made at the end of this period. the mic for terbinafine ranged from 0.055μg/ml to 0.109μg/ml, and the mic for itraconazole ranged from 0.219μg/ml to 1.75μg/ml. for both drugs, the mic from the isolates from ioc was 0.875μg/ml. the present study demonstrates the high susceptibility of sporothrix schenckii to terbinafine. further studies to correlate the in vitro susceptibility tests with the clinical response of patients with sporotrichosis are needed.
Molecular characterisation of Sporothrix schenckii isolates from humans and cats involved in the sporotrichosis epidemic in Rio de Janeiro, Brazil
Reis, Rosani Santos;Almeida-Paes, Rodrigo;Muniz, Mauro de Medeiros;Tavares, Patrícia Morais e Silva;Monteiro, Paulo Cezar Fialho;Schubach, Tania Maria Pacheco;Gutierrez-Galhardo, Maria Clara;Zancopé-Oliveira, Rosely Maria;
Memórias do Instituto Oswaldo Cruz , 2009, DOI: 10.1590/S0074-02762009000500018
Abstract: an epidemic of sporotrichosis, a subcutaneous mycosis caused by the fungus sporothrix schenckii, is ongoing in rio de janeiro, brazil, in which cases of human infection are related to exposure to cats. in an attempt to demonstrate the zoonotic character of this epidemic using molecular methodology, we characterised by dna-based typing methods 19 human and 25 animal s. schenckii isolates from the epidemic, as well as two control strains. to analyse the isolates, the random amplified polymorphic dna (rapd) technique was performed using three different primers, together with dna fingerprinting using the minisatellite derived from the wild-type phage m13 core-sequence. the analyses generated amplicons with considerable polymorphism. although isolates exhibited high levels of genetic relatedness, they could be clustered into 5-10 genotypes. the rapd profiles of epidemic s. schenckii isolates could be distinguished from that of the united states isolate, displaying 20% similarity to each primer and 60% when amplified with the m13 primer. dna fingerprinting of s. schenckii isolated from the nails (42.8%) and the oral cavities (66%) of cats were identical to related human samples, suggesting that there is a common infection source for animals and humans in this epidemic. it is clear that cats act as a vehicle for dissemination of s. schenckii.
Aislamiento ambiental de Sporothrix globosa en relación a un caso de esporotricosis linfo-cutánea Sporothrix globosa isolation related to a case of lymphocutaneous sporotrichosis  [cached]
Rodrigo Cruz,Peggy Vieille,David Oschilewski
Revista chilena de infectología , 2012,
Abstract: Introducción: Sporothrix schenckii complex comprende un grupo de hongos dimórficos ambientales que provocan esporotricosis. En Chile se han comunicado casos aislados en humanos; sin embargo, no se han descrito aislamientos de este hongo desde el ambiente. Objetivos: Orientados por un caso de esporotricosis linfo-cutánea en una paciente de 75 a os, se realizó un estudio ambiental para lograr el aislamiento de S. complex desde el suelo donde la paciente realizaba labores de horticultura. Materiales y Métodos: Se extrajo tierra y restos vegetales de cinco sectores donde la paciente efectuaba sus labores de horticultura. Se realizaron diluciones de tierras y siembra en agar Sabouraud con cicloheximida y cloranfenicol a 26°C. Los restos vegetales fueron sembrados directamente en el mismo medio. Las colonias sugerentes de S. complex se resembraron en agar PDA a 26°C y se identificaron según recomendación de Marimon y cols. Resultados: De las diez placas del primer muestreo, una colonia de la siembra de tierra se identificó como Sporothrix globosa. En el segundo muestreo hubo desarrollo de S. globosa en dos placas sembradas con tierra, con un total de seis colonias. No hubo desarrollo de S. complex en los restos vegetales. La cepa aislada desde la paciente también fue identificada como S. globosa. Conclusión: Por primera vez en Chile se logra el aislamiento del ambiente de una especie de S. complex. Sporothrix globosa fue la especie identificada tanto del suelo como desde la paciente con esporotricosis. Introduction: Sporothrix schenckii complex comprises a group of environmental dimorphic fungi that cause sporotrichosis. In Chile, isolated cases have been reported in humans, though no environmental isolates have been described. Objective: To achieve isolation of Sporothrix complex from the soil where a 75 year old patient with lymphocutaneous sporotrichosis performs horticulture work. Materials and Methods: In March and July 2011 soil and plant debris from five sectors where the patient does his work in horticulture was extracted. The soil samples were diluted and inoculated in Sabouraud agar with cycloheximide and chloramphenicol at 26 °C. The plant debris was directly inoculated in the same medium. Colonies suggestive of Sporothrix complex were reseeded in PDA agar at 26 ° C and identified as recommended by Marimon et al. Results: Of the 10 plates from the first sampling, one colony was identified as Sporothrix globosa. In the second sampling, Sporothrix globosa grew in two plates seeded with soil, with a total of 6 colonies. There was no growth of Sporothr
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