scholarly journals Identification of a gene encoding adaptin-like protein in the Paracoccidioides brasiliensis genome by random amplified polymorphic DNA analysis

2007 ◽  
Vol 56 (7) ◽  
pp. 884-887 ◽  
Author(s):  
Patrícia Ferrari Andreotti ◽  
Juliana Leal Monteiro da Silva ◽  
Elaine Cristina Teixeira ◽  
Maria Célia Bertolini ◽  
Christiane Pienna Soares ◽  
...  

Paracoccidioides brasiliensis isolates are not homogeneous in their patterns of pathogenicity in animals and adhesion to epithelial cells. During this investigation, genotypic differences were observed between two samples of P. brasiliensis strain 18 yeast phase (Pb18) previously cultured many times, one taken before (Pb18a) and the other after (Pb18b) animal inoculation. Random amplified polymorphic DNA analysis using the primer OPJ4 distinguished Pb18b from Pb18a by one 308 bp DNA fragment, which after cloning and sequencing was shown to encode a polypeptide sequence homologous to the protein β-adaptin. It is suggested, by comparison to other micro-organisms, that this protein might play an important role in the virulence of P. brasiliensis. This result demonstrates the influence of in vitro subculturing on the genotype of this organism.

2005 ◽  
Vol 47 (3) ◽  
pp. 119-123 ◽  
Author(s):  
Sarah Desirée Barbosa Cavalcanti ◽  
José Eduardo Levi ◽  
Kátia Cristina Dantas ◽  
José Eduardo Costa Martins

Our purpose was to compare the genetic polymorphism of six samples of P. brasiliensis (113, 339, BAT, T1F1, T3B6, T5LN1), with four samples of P. cerebriformis (735, 741, 750, 361) from the Mycological Laboratory of the Instituto de Medicina Tropical de São Paulo, using Random Amplified Polymorphic DNA Analysis (RAPD). RAPD profiles clearly segregated P. brasiliensis and P. cerebriformis isolates. However, the variation on band patterns among P. cerebriformis isolates was high. Sequencing of the 28S rDNA gene showed nucleotide conservancy among P. cerebriformis isolates, providing basis for taxonomical grouping, and disclosing high divergence to P. brasiliensis supporting that they are in fact two distinct species. Moreover, DNA sequence suggests that P. cerebriformis belongs in fact to the Aspergillus genus.


2020 ◽  
Vol 4 (2) ◽  
pp. 57-62
Author(s):  
IRFAN MARTIANSYAH ◽  
NURHAIMI HARIS ◽  
TATI HUSNIYATI ◽  
EDI DJAUHARI PURWAKUSUMAH

The rubber seeds are insufficient for producing rootstocks to rubber grafting. It can be overcome by an in vitro micro-cutting culture technique developed in the Indonesian Research Institute for Biotechnology and Bioindustry (IRIBB). However, the origin clone of 57 rubber genotypes used as an explant source in vitro micro-cutting culture is not recognized. The study was to investigate the 57 genotypes that came from mixed GT 1, PB 260, and RRIM 600 as parent clones. We investigated using seven primers of Random Amplified Polymorphic DNA (RAPD), i.e., OPA 02, OPA 07, OPA 15, OPB 04, OPC 05, OPC 11, and OPC 20. The qualitative analyzed by electrophoresis 1% gel agarose. A total of 47 DNA fragments produced with an average of 7 fragments per primer. OPA 02 generated of 13 fragments, whereas OPB 04 only one fragment. The DNA fragment pattern shows the presence of polymorphism. The genetic similarity coefficients obtained in the range of 62-96%. The highest genetic similarity (96%) is genotype 70 and 78. It recognized that 42 genotypes from 57 rubber genotypes had the closest relationship with PB 260 clones. Furthermore, six genotypes had a significant growth response as an explant in vitro micro-cutting culture.


1998 ◽  
Vol 42 (11) ◽  
pp. 2836-2840 ◽  
Author(s):  
Francis Mégraud ◽  
Alessandra Occhialini ◽  
Jean François Rossignol

ABSTRACT Nitazoxanide, a thiazolide compound, and its desacetyl derivative, tizoxanide, have antimicrobial properties against anaerobic bacteria, as well as against helminths and protozoa. Because the treatment ofHelicobacter pylori infection may be jeopardized by metronidazole resistance, nitazoxanide and tizoxanide were tested in vitro against these bacteria. The MICs of these two compounds were determined by agar dilution and were compared to those of metronidazole. Exposure to subinhibitory concentrations of nitazoxanide was also carried out by the method of Szybalski (W. Szybalski and V. Bryson, J. Bacteriol. 64:489–499, 1952). The MICs of nitazoxanide and tizoxanide for 103 strains ranged from 0.25 to 8 μg/ml, with the MIC at which 50% of strains are inhibited (MIC50) being 1 μg/ml and the MIC90 being 4 μg/ml, and no resistant strain was detected, whereas strains resistant to metronidazole were detected. When 10 strains were successively subcultured on medium containing nitazoxanide, no significant change in the MICs of this compound was observed. A pilot study of nitazoxanide for the treatment of H. pyloriinfection was carried out with 86 patients in association with 20 mg of omeprazole. An eradication rate of 83% (95% confidence interval, 64% to 94%) was obtained in a per-protocol analysis in the group receiving 1 g of nitazoxanide orally twice daily, and a few side effects were observed. The failures could not be explained by the selection of resistant strains since the MICs of nitazoxanide were similar for six pairs of isolates (proven to be the same strain by random amplified polymorphic DNA analysis in four cases) cultured before and after the treatment failure. Nitazoxanide exhibits good antimicrobial activity against H. pylori without the problem of acquired resistance which is encountered with metronidazole and has been demonstrated to have a satisfactory effect in a dose-ranging pilot study. It is therefore a good candidate to be included in treatment regimens aimed at the eradication of H. pylori.


2005 ◽  
Vol 47 (5) ◽  
pp. 257-262 ◽  
Author(s):  
Cilmery S. Kurokawa ◽  
Catalina R. Lopes ◽  
Maria F. Sugizaki ◽  
Eiko E. Kuramae ◽  
Marcello F. Franco ◽  
...  

Ten isolates of Paracoccidioides brasiliensis were examined for differences in virulence in outbred mice intravenously inoculated with the fungus, associated with mycelial morphology, and genetic patterns measured by random amplified polymorphic DNA (RAPD). Virulence was evaluated by viable yeast cell recovery from lungs and demonstration of histopathologic lesions in different organs. The results showed that the isolates presented four virulence degrees: high virulence, intermediate, low and non-virulence. RAPD clustered the isolates studied in two main groups with 56% of genetic similarity. Strains with low virulence, Pb265 or the non-virulent, Pb192, showed glabrous/cerebriform morphology and high genetic similarity (98.7%) when compared to the other isolates studied. The same was observed with Bt79 and Bt83 that shared 96% genetic similarity, cottony colonies and high virulence. The RAPD technique could only discriminate P. brasiliensis isolates according to glabrous/cerebriform or cottony colonies, and also high from low virulence strains. Isolates with intermediate virulence such as Pb18, Pb18B6, Bt32 and Bt56 showed variability in their similarity coefficient suggesting that RAPD was able to detect genetic variability in this fungal specie. Virulence profile of P. brasiliensis demonstrated that both mycelial morphologic extreme phenotypes may be associated with fungal virulence and their in vitro subculture time. Thus, RAPD technique analysis employed in association with virulence, morphologic and immunologic aspects might prove adequate to detect differences between P. brasiliensis isolates.


1995 ◽  
Vol 33 (2) ◽  
pp. 505-507 ◽  
Author(s):  
C M Soares ◽  
E E Madlun ◽  
S P da Silva ◽  
M Pereira ◽  
M S Felipe

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