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Search Results: 1 - 10 of 112892 matches for " Gusm?o Leonor "
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Rapid identification of Aspergillus fumigatus within the section Fumigati
Rita Serrano, Leonor Gusmo, António Amorim, Ricardo Araujo
BMC Microbiology , 2011, DOI: 10.1186/1471-2180-11-82
Abstract: A multiplex PCR was developed using prior information based on β-tubulin (βtub) and rodlet A (rodA) partial gene sequences. PCR amplification of βtub and rodA fragments resulted in a distinctive electrophoretic pattern in A. fumigatus and N. udagawae. The polymorphisms found in the smallest amplified sequence of βtub (153 bp) and rodA (103 bp) genes were then compared among and within species of this taxonomic section. βtub was able to differentiate among 13 individual species and two groups of species that included the pathogenic fungus A. lentulus. A more limited number of sequences were available for rodA; nevertheless, we were able to distinguish Aspergillus viridinutans, N. hiratsukae and N. udagawae.The assay described in the present study proved to be specific and highly reproducible, representing a fast and economic way of targeting molecular identification of the relevant mould, A. fumigatus, in clinical laboratories.Aspergillosis is the most common invasive mould disease worldwide. Recently, molecular techniques have been applied to fungal diagnosis and to the identification of species, and new fungal species that are morphologically similar to A. fumigatus have been described, authenticated and included in section Fumigati [1-3]. Therefore, this section now includes a few anamorphous Aspergillus species and teleomorphic species that are found in the genus Neosartorya [4]. The characteristics of the colonies on standard culture media are often similar to A. fumigatus, but conidia may be rather distinct. Neosartorya species produce heat-resistant ascospores [4].Misidentification of fungal species within the section Fumigati has been increasingly reported by clinical laboratories. Species, such as Aspergillus lentulus, Aspergillus viridinutans, Aspergillus fumigatiaffinis, Aspergillus fumisynnematus, Neosartorya pseudofischeri, Neosartorya hiratsukae and Neosartorya udagawae, are frequently reported as A. fumigatus [1,2,5,6]. Some of these species have been
Diversity and specificity of microsatellites within Aspergillus section Fumigati
Araujo Ricardo,Amorim António,Gusmo Leonor
BMC Microbiology , 2012, DOI: 10.1186/1471-2180-12-154
Abstract: Background Microsatellites (or short tandem repeats, STRs) are the genetic markers of choice for studying Aspergillus fumigatus molecular epidemiology due to its reproducibility and high discrimination power. However, the specificity of these markers must be investigated in a group of isolates from closely related species. The aim of this work was to test a microsatellite-based PCR multiplex previously designed for A. fumigatus in a set of species belonging to section Fumigati, namely Aspergillus fumigatiaffinis, Aspergillus lentulus, Aspergillus novofumigatus, Aspergillus unilateralis, Aspergillus viridinutans, Neosartorya fischeri, Neosartorya hiratsukae, Neosartorya pseudofischeri and Neosartorya udagawae. Results The reference A. fumigatus strain ATCC 46645 was easily genotyped in standard conditions showing a final electrophoretic profile of 8 expected peaks corresponding to each microsatellite locus. Inversely, no peaks were observed for all other species from section Fumigati, with an exception for marker MC6b in A. unilateralis. By screening the genome sequence of Neosartorya fischeri NRRL 181, the results showed that MC3, MC6a and MC7 might be employed for N. fischeri genotyping since these markers present several repeats of each motif. The accumulation of insertions and deletions was frequently observed in the genomic regions surrounding the microsatellites, including those where the A. fumigatus primers are located. The amplification of microsatellite markers in less stringent amplification conditions resulted in a distinct electrophoretic profile for species within section Fumigati. Conclusions Therefore, the microsatellite-based PCR multiplex allow simple identification of A. fumigatus and, with a slight modification of temperature conditions, it also allows discriminating other pathogenic species within section Fumigati, particularly A. fumigatiaffinis, N. fischeri and N. udagawae.
Novos registros de Hyphomycetes decompositores para o Estado da Bahia, Brasil
Barbosa, Flávia Rodrigues;Maia, Leonor Costa;Gusmo, Luís Fernando Pascholati;
Acta Botanica Brasilica , 2009, DOI: 10.1590/S0102-33062009000200004
Abstract: leaf litter from clusia melchiorii gleason and c. nemorosa g. mey. was collected bimonthly at serra da jibóia, state of bahia, from october/2005 to june/2006. the leaves were washed with tap water and maintained in a moist chamber for 30 days. the fungal structures were collected and mounted in permanent slides with pvl. descriptions and illustrations of seven new records of hyphomycetes from bahia state are presented [beltrania querna harkn., clonostachys compactiuscula (sacc.) d. hawksw. & w. gams, dictyosporium elegans corda, gyrothrix verticiclada (goid.) s. hughes & piroz., pseudobotrytis terrestris (timonin) subram., sporendocladia bactrospora (w.b. kendr.) m.j. wingf. and stachybotrys parvispora s. hughes].
Fungos conidiais associados ao folhedo de Clusia melchiorii Gleason e C. nemorosa G. Mey. (Clusiaceae) em fragmento de Mata Atlantica, BA, Brasil
Barbosa, Flávia Rodrigues;Maia, Leonor Costa;Gusmo, Luís Fernando Pascholati;
Acta Botanica Brasilica , 2009, DOI: 10.1590/S0102-33062009000100010
Abstract: in order to increase the diversity knowledge of conidial fungi, 10 dead leaves from three individuals of c. nemorosa and c. melchiorii were bimonthly collected at the "serra da jibóia", state of bahia, from october/2005 to june/2006. the leaves were washed with tap water and maintained in moist chamber during 30 days. the fungal structures were collected for morphological studies. seventy nine taxa of ascomycota, in the anamorphic state were registered: 78 hyphomycete and one coelomycete. most of the species occurred on clusia melchiorii (87%) and 55% on c. nemorosa. the majority of the fungi presented sporadical frequency and accidental constancy. the most frequent species were: beltrania rhombica penz., chaetopsina fulva rambelli, dactylaria ficusicola paulus, gadek & hyde, verticillium theobromae (turconi) mason & hughes e volutella sp. 1 (on c. melchiorii) and atroseptaphiale flagelliformis matsush., pseudobeltrania sp., zygosporium gibbum (sacc., rousseau & bommer) hughes, verticillium theobromae (turconi) mason & hughes and volutella sp. 1 (on c. nemorosa). the similarity of fungi between the two species of clusia reached 60% and 11 taxa were constant in both hosts: atrosetaphiale flagelliformis, beltraniella portoricensis (stevens) piroz. & patil, chalara alabamensisjones & ingram., cryptophiale kakombensis piroz., parasympodiella laxa (subram. & vittal), speiropsis scopiformis kuthub. & nawawi, thozetella cristata piroz. & hodges, umbellidion radulans sutton & hodges, verticillium theobromae, volutella sp. 2 and zygosporium gibbum. the data show that the litter produced by c. melchiorii and c. nemorosa, at the serra da jibóia, is rich in conidial fungi. these fungi, as decomposers, are important for the dinamic of the studied ecosystem.
Riqueza de espécies de fungos conidiais em duas áreas de Mata Atlantica no Morro da Pioneira, Serra da Jibóia, BA, Brasil
Marques, Marcos Fabio Oliveira;Gusmo, Luis Fernando Pascholati;Maia, Leonor Costa;
Acta Botanica Brasilica , 2008, DOI: 10.1590/S0102-33062008000400006
Abstract: the fragment of atlantic forest at serra da jibóia, municipality of santa terezinha, bahia state, is a priority area for conservation. five expeditions were undertaken every two months, from october/2005 to june/2006, in order to investigate the fungi that decompose leaf litter in this ecosystem. thirty samples of plant debris (leaves, petioles, twigs and bark) were collected in three parcels of 10 m2, 10 m from each other, in two areas with different vegetation and humidity. samples were washed in tap water and incubated in moist chambers for 30 days; during this period fungal structures on substrates were studied and 106 species of conidial fungi were identified. in spite of the similar number of species, there was low similarity (25%) between the communities of conidial fungi of the two areas according to s?rensen's index. these data increase our knowledge regarding the distribution and diversity of conidial fungi that colonize plant debris in the atlantic forest and confirm species richness in the areas studied.
Espécies de Vermiculariopsiella (Hyphomycetes) associadas a substratos vegetais em fragmento de Mata Atlantica, Serra da Jibóia, Estado da Bahia, Brasil
Marques, Marcos Fabio Oliveira;Gusmo, Luis Fernando Pascholati;Maia, Leonor Costa;
Brazilian Journal of Botany , 2008, DOI: 10.1590/S0100-84042008000400011
Abstract: during a survey of microfungi associated to plant debris in a fragment of atlantic rain forest, serra da jibóia, santa terezinha, bahia state, from october/2005 to june/2006, four vermiculariopsiella species were found in association to the decomposition of leaves, petioles and twigs in this ecosystem. v. immersa (desm.) bender and v. cubensis (r.f. casta?eda) nawawi, kuthub. & b. sutton are new records for "serra da jibóia" and state of bahia, respectively, v. cornuta (v. rao & hoog) nawawi, kuthub. & b. sutton and v. falcata nawawi, kuthub. & b. sutton represent new records for south america. descriptions and illustrations of morphologic characteristics of the four species, information about substrates and geographical distribution as well as a taxonomic key for all known species of vermiculariopsiella are provided.
Análisis de la estructura genética en una muestra poblacional de Bucaramanga, departamento de Santander
Martha Lucía Hincapié,Adriana María Gil,Adriana Lucía Pico,Leonor Gusmo
Colombia Médica , 2009,
Abstract: Introducción: El fenómeno de sub-estructura en las poblaciones ha tenido desde hace varios a os un abordaje amplio, que se enfocó, entre otros, en la identificación y cuantificación de la mezcla étnica presente en estudios de mapeo asociativo, para comprobar la asociación de marcadores polimórficos en el desarrollo de enfermedades comunes complejas, como responsable de falsos positivos. No obstante el reconocimiento de este problema, no se tiene suficiente información genética en el contexto nacional ni local que permita determinar la posible diferenciación de subgrupos poblacionales en cada región en particular.Objetivo: Determinar la estructura genética en una muestra poblacional de la ciudad de Bucaramanga, a partir del análisis de 19 marcadores microsatélites autosómicos en distintos subgrupos poblacionales.Metodología: De la base de datos del Laboratorio de Genética Humana de la Universidad Industrial de Santander, se seleccionaron aleatoriamente 350 muestras de ADN, y se amplificaron 19 marcadores autosómicos Short Tandem Repeat mediante los kits Powerplex 16 y FFFL (Promega) .Resultados: En el análisis de equilibrio Hardy Weinberg, no se obtuvieron diferencias estadísticamente significativas en 18 de 19 marcadores Short Tandem Repeat autosómicos analizados en la población de Bucaramanga. El único marcador que mostró no estar en equilibrio Hardy Weinberg en la población de Bucaramanga fue el F13B (valor de significancia de p=0.00264, después de aplicar la corrección de Bonferroni).Discusión: Las poblaciones representadas en los seis estratos socioeconómicos mostraron alta diversidad genética intragrupos, que ratificó una alta variabilidad entre los individuos de la ciudad de Bucaramanga, acorde con el bajo valor de FST entre distintos grupos, determinado en el análisis molecular de varianza con base en frecuencias alélicas observadas para los 19 Short Tandem Repeat analizados.Conclusión: La alta diversidad genética y el análisis molecular de varianza mostraron una baja diferenciación entre los seis estratos socioeconómicos en la ciudad de Bucaramanga, y evidenciaron a la población como una misma unidad genética, no sub-estructurada.
SNaPAfu: A Novel Single Nucleotide Polymorphism Multiplex Assay for Aspergillus fumigatus Direct Detection, Identification and Genotyping in Clinical Specimens
Rita Caramalho, Leonor Gusmo, Michaela Lackner, António Amorim, Ricardo Araujo
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0075968
Abstract: Objective Early diagnosis of invasive aspergillosis is essential for positive patient outcome. Likewise genotyping of fungal isolates is desirable for outbreak control in clinical setting. We designed a molecular assay that combines detection, identification, and genotyping of Aspergillus fumigatus in a single reaction. Methods To this aim we combined 20 markers in a multiplex reaction and the results were seen following mini-sequencing readings. Pure culture extracts were firstly tested. Thereafter, Aspergillus-DNA samples obtained from clinical specimens of patients with possible, probable, or proven aspergillosis according to European Organization for the Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) criteria. Results A new set of designed primers allowed multilocus sequence typing (MLST) gene amplification in a single multiplex reaction. The newly proposed SNaPAfu assay had a specificity of 100%, a sensitivity of 89% and detection limit of 1 ITS copy/mL (~0.5 fg genomic Aspergillus-DNA/mL). The marker A49_F was detected in 89% of clinical samples. The SNaPAfu assay was accurately performed on clinical specimens using only 1% of DNA extract (total volume 50 μL) from 1 mL of used bronchoalveolar lavage. Conclusions The first highly sensitive and specific, time- and cost-economic multiplex assay was implemented that allows detection, identification, and genotyping of A. fumigatus strains in a single amplification followed by mini-sequencing reaction. The new test is suitable to clinical routine and will improve patient management.
Analysis of paternal lineages in Brazilian and African populations
Carvalho, Mónica;Brito, Pedro;Lopes, Virgínia;Andrade, Lisa;Anjos, Ma Jo?o;Real, Francisco Corte;Gusmo, Leonor;
Genetics and Molecular Biology , 2010, DOI: 10.1590/S1415-47572010005000067
Abstract: the present-day brazilian population is a consequence of the admixture of various peoples of very different origins, namely, amerindians, europeans and africans. the proportion of each genetic contribution is known to be very heterogeneous throughout the country. the aim of the present study was to compare the male lineages present in two distinct brazilian populations, as well as to evaluate the african contribution to their male genetic substrate. thus, two brazilian population samples from manaus (state of amazon) and ribeir?o preto (state of s?o paulo) and three african samples from guinea bissau, angola and mozambique were typed for a set of nine y chromosome specific strs. the data were compared with those from african, amerindian and european populations. by using y-str haplotype information, low genetic distances were found between the manaus and ribeir?o preto populations, as well as between these and others from iberia. likewise, no significant distances were observed between any of the african samples from angola, mozambique and guinea bissau. highly significant rst values were found between both brazilian samples and all the african and amerindian populations. the absence of a significant sub-saharan african male component resulting from the slave trade, and the low frequency in amerindian ancestry y-lineages in the manaus and ribeir?o preto population samples are in accordance with the accentuated gender asymmetry in admixture processes that has been systematically reported in colonial south american populations.
SNaPaer: A Practical Single Nucleotide Polymorphism Multiplex Assay for Genotyping of Pseudomonas aeruginosa
Nadia Eusebio, Tiago Pinheiro, Adelina A. Amorim, Fernanda Gamboa, Lucília Saraiva, Leonor Gusmo, António Amorim, Ricardo Araujo
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0066083
Abstract: Multilocus sequence typing (MLST) represents the gold standard genotyping method in studies concerning microbial population structure, being particularly helpful in the detection of clonal relatedness. However, its applicability on large-scale genotyping is limited due to the high cost and time spent on the task. The selection of the most informative nucleotide positions simplifies genomic characterization of bacteria. A simple and informative multiplex, SNaPaer assay, was developed and genotyping of Pseudomonas aeruginosa was obtained after a single reaction of multiplex PCR amplification and mini-sequencing. This cost-effective technique allowed the analysis of a Portuguese set of isolates (n = 111) collected from three distinct hospitals and the genotyping data could be obtained in less than six hours. Point mutations were shown to be the most frequent event responsible for diversification of the Portuguese population sample. The Portuguese isolates corroborated the epidemic hypothesis for P. aeruginosa population. SNaPaer genotyping assay provided a discriminatory power of 0.9993 for P. aeruginosa, by testing in silico several hundreds of MLST profiles available online. The newly proposed assay targets less than 0.01% of the total MLST length and guarantees reproducibility, unambiguous analysis and the possibility of comparing and transferring data between different laboratories. The plasticity of the method still supports the addition of extra molecular markers targeting specific purposes/populations. SNaPaer can be of great value to clinical laboratories by facilitating routine genotyping of P. aeruginosa.
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