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21.
Letícia Muraro Wildner Maria Luiza Bazzo Susie Coutinho Liedke Christiane Louren?o Nogueira Gabriela Segat Simone Gon?alves Senna Aline Daiane Schlindwein Jaquelline Germano de Oliveira Darcita B Rovaris Claudio A Bonjardim Erna G Kroon Paulo CP Ferreira 《Memórias do Instituto Oswaldo Cruz》2014,109(3):356-361
The identification of mycobacteria is essential because tuberculosis (TB) and
mycobacteriosis are clinically indistinguishable and require different therapeutic
regimens. The traditional phenotypic method is time consuming and may last up to 60
days. Indeed, rapid, affordable, specific and easy-to-perform identification methods
are needed. We have previously described a polymerase chain reaction-based method
called a mycobacteria mobility shift assay (MMSA) that was designed for
Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria
(NTM) species identification. The aim of this study was to assess the MMSA for the
identification of MTC and NTM clinical isolates and to compare its performance with
that of the PRA-hsp65 method. A total of 204 clinical isolates (102
NTM and 102 MTC) were identified by the MMSA and PRA-hsp65. For
isolates for which these methods gave discordant results, definitive species
identification was obtained by sequencing fragments of the 16S rRNA and
hsp65 genes. Both methods correctly identified all MTC isolates. Among
the NTM isolates, the MMSA alone assigned 94 (92.2%) to a complex or species, whereas
the PRA-hsp65 method assigned 100% to a species. A 91.5% agreement
was observed for the 94 NTM isolates identified by both methods. The MMSA provided
correct identification for 96.8% of the NTM isolates compared with 94.7% for
PRA-hsp65. The MMSA is a suitable auxiliary method for routine
use for the rapid identification of mycobacteria. 相似文献
22.
Microsporidia, amitochondrial protists, possess a 70-kDa heat shock protein gene of mitochondrial evolutionary origin 总被引:13,自引:3,他引:10
Peyretaillade E; Broussolle V; Peyret P; Metenier G; Gouy M; Vivares CP 《Molecular biology and evolution》1998,15(6):683-689
An intronless gene encoding a protein of 592 amino acid residues with
similarity to 70-kDa heat shock proteins (HSP70s) has been cloned and
sequenced from the amitochondrial protist Encephalitozoon cuniculi (phylum
Microsporidia). Southern blot analyses show the presence of a single gene
copy located on chromosome XI. The encoded protein exhibits an N-terminal
hydrophobic leader sequence and two motifs shared by proteobacterial and
mitochondrially expressed HSP70 homologs. Phylogenetic analysis using
maximum likelihood and evolutionary distances place the E. cuniculi
sequence in the cluster of mitochondrially expressed HSP70s, with a higher
evolutionary rate than those of homologous sequences. Similar results were
obtained after cloning a fragment of the homologous gene in the closely
related species E. hellem. The presence of a nuclear targeting signal-like
sequence supports a role of the Encephalitozoon HSP70 as a molecular
chaperone of nuclear proteins. No evidence for cytosolic or endoplasmic
reticulum forms of HSP70 was obtained through PCR amplification. These data
suggest that Encephalitozoon species have evolved from an ancestor bearing
mitochondria, which is in disagreement with the postulated presymbiotic
origin of Microsporidia. The specific role and intracellular localization
of the mitochondrial HSP70-like protein remain to be elucidated.
相似文献
23.
24.
25.
Lucas A Smolders Bj?rn P Meij David Onis Frank M Riemers Niklas Bergknut Richard Wubbolts Guy CM Grinwis Martin Houweling Marian JA Groot Koerkamp Dik van Leenen Frank CP Holstege Herman AW Hazewinkel Laura B Creemers Louis C Penning Marianna A Tryfonidou 《Arthritis research & therapy》2013,15(1):R23
Introduction
Early degeneration of the intervertebral disc (IVD) involves a change in cellular differentiation from notochordal cells (NCs) in the nucleus pulposus (NP) to chondrocyte-like cells (CLCs). The purpose of this study was to investigate the gene expression profiles involved in this process using NP tissue from non-chondrodystrophic and chondrodystrophic dogs, a species with naturally occurring IVD degeneration.Methods
Dual channel DNA microarrays were used to compare 1) healthy NP tissue containing only NCs (NC-rich), 2) NP tissue with a mixed population of NCs and CLCs (Mixed), and 3) NP tissue containing solely CLCs (CLC-rich) in both non-chondrodystrophic and chondrodystrophic dogs. Based on previous reports and the findings of the microarray analyses, canonical Wnt signaling was further evaluated using qPCR of relevant Wnt target genes. We hypothesized that caveolin-1, a regulator of Wnt signaling that showed significant changes in gene expression in the microarray analyses, played a significant role in early IVD degeneration. Caveolin-1 expression was investigated in IVD tissue sections and in cultured NCs. To investigate the significance of Caveolin-1 in IVD health and degeneration, the NP of 3-month-old Caveolin-1 knock-out mice was histopathologically evaluated and compared with the NP of wild-type mice of the same age.Results
Early IVD degeneration involved significant changes in numerous pathways, including Wnt/β-catenin signaling. With regard to Wnt/β-catenin signaling, axin2 gene expression was significantly higher in chondrodystrophic dogs compared with non-chondrodystrophic dogs. IVD degeneration involved significant down-regulation of axin2 gene expression. IVD degeneration involved significant down-regulation in Caveolin-1 gene and protein expression. NCs showed abundant caveolin-1 expression in vivo and in vitro, whereas CLCs did not. The NP of wild-type mice was rich in viable NCs, whereas the NP of Caveolin-1 knock-out mice contained chondroid-like matrix with mainly apoptotic, small, rounded cells.Conclusions
Early IVD degeneration involves down-regulation of canonical Wnt signaling and Caveolin-1 expression, which appears to be essential to the physiology and preservation of NCs. Therefore, Caveolin-1 may be regarded an exciting target for developing strategies for IVD regeneration. 相似文献26.
C. P. Larralde-Corona P. C. González-Blanco G. Viniegra-González 《Biotechnology Techniques》1994,8(4):261-266
Summary A new equation is proposed in order to obtain a consistent and accurate correlation between morphometric data of growing mycelia of Gibberella fujikuroi in solid media and its observed specific growth rates (
obs) in stirred fermenters. A critical analysis is made of previous data and models. 相似文献
27.
Seed dispersal plays a critical role in rainforest regeneration patterns, hence loss of avian seed dispersers in fragmented landscapes may disrupt forest regeneration dynamics. To predict whether or not a plant will be dispersed in fragmented forests, it is necessary to have information about frugivorous bird distribution and dietary composition. However, specific dietary information for frugivorous birds is often limited. In such cases, information on the seed-crushing behaviour, gape width and relative dietary dominance by fruit may be used to describe functional groups of bird species with respect to their potential to disperse similar seeds. We used this information to assess differences in the seed dispersal potential of frugivorous bird assemblages in a fragmented rainforest landscape of southeast Queensland, Australia. The relative abundance of frugivorous birds was surveyed in extensive, remnant and regrowth rainforest sites (16 replicates of each). Large-gaped birds with mixed diets and medium-gaped birds with fruit-dominated diets were usually less abundant in remnants and regrowth than in continuous forest. Small-gaped birds with mixed diets and birds with fruit as a minor dietary component were most abundant in regrowth. We recorded a similar number of seed-crushing birds and large-gaped birds with fruit-dominated diets across site types. Bird species that may have the greatest potential to disperse a large volume and wide variety of plants, including large-seeded plants, tended to be less abundant outside of extensive forests, although one species, the figbird Sphecotheres viridis, was much more abundant in these areas. The results suggest that the dispersal of certain plant taxa would be limited in this fragmented landscape, although the potential for the dispersal of large-seeded plants may remain, despite the loss of several large-gaped disperser species. 相似文献
28.
29.
A simple and accurate method for determining the distribution of sizes of single cells and aggregates of Azotobacter vinelandii by image analysis has been developed. A staining procedure using methylene blue helps to enhance the contrast between aggregates and background without altering aggregate size distribution. Sample dilution affected the distribution of the population and therefore should be avoided. Mixing and aeration conditions during culture play an important role in the aggregation of A. vinelandii. Cells grown under mild mixing conditions (unbaffled flasks) presented a thick slime layer and formed aggregates of up to 35 microm of average equivalent diameter. In contrast, under strong agitation conditions (baffled flasks) practically no aggregates were formed throughout cultivation. The method described can be used for the characterization of aggregation of other microbial cultures. 相似文献
30.