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1.
Microbiol Spectr ; 10(6): e0146622, 2022 12 21.
Artigo em Inglês | MEDLINE | ID: mdl-36255300

RESUMO

Newborn screening for cystic fibrosis (CF) can identify affected but asymptomatic infants. The selection of omic technique for gut microbiota study is crucial due to both the small amount of feces available and the low microorganism load. Our aims were to compare the agreement between 16S rRNA amplicon sequencing and metaproteomics by a robust statistical analysis, including both presence and abundance of taxa, to describe the sequential establishment of the gut microbiota during the first year of life in a small size sample (8 infants and 28 fecal samples). The taxonomic assignations by the two techniques were similar, whereas certain discrepancies were observed in the abundance detection, mostly the lower predicted relative abundance of Bifidobacterium and the higher predicted relative abundance of certain Firmicutes and Proteobacteria by amplicon sequencing. During the first months of life, the CF gut microbiota is characterized by a significant enrichment of Ruminococcus gnavus, the expression of certain virulent bacterial traits, and the detection of human inflammation-related proteins. Metaproteomics provides information on composition and functionality, as well as data on host-microbiome interactions. Its strength is the identification and quantification of Actinobacteria and certain classes of Firmicutes, but alpha diversity indices are not comparable to those of amplicon sequencing. Both techniques detected an aberrant microbiota in our small cohort of infants with CF during their first year of life, dominated by the enrichment of R. gnavus within a human inflammatory environment. IMPORTANCE In recent years, some techniques have been incorporated for the study of microbial ecosystems, being 16S rRNA gene sequencing being the most widely used. Metaproteomics provides the advantage of identifying the interaction between microorganisms and human cells, but the available databases are less extensive as well as imprecise. Few studies compare the statistical differences between the two techniques to define the composition of an ecosystem. Our work shows that the two methods are comparable in terms of microorganism identification but provide different results in alpha diversity analysis. On the other hand, we have studied newborns with cystic fibrosis, for whom we have described the establishment of an intestinal ecosystem marked by the inflammatory response of the host and the enrichment of Ruminococcus gnavus.


Assuntos
Fibrose Cística , Microbioma Gastrointestinal , Microbiota , Humanos , Recém-Nascido , Lactente , RNA Ribossômico 16S/genética , Microbioma Gastrointestinal/genética , Fibrose Cística/microbiologia , Bactérias , Fezes/microbiologia , Firmicutes/genética , Microbiota/genética
2.
Int J Parasitol ; 50(5): 377-388, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32360428

RESUMO

The biological variability among Neospora caninum isolates has been widely shown, however, the molecular basis that determines this diversity has not been thoroughly elucidated to date. The latest studies have focused on a limited number of isolates. Therefore, the goal of the present study was to compare the proteome of a larger number of N. caninum isolates with different origins and virulence. Label-free LC-MS/MS was used to investigate the tachyzoite proteomic differences among Nc-Bahia, Nc-Spain4H and Nc-Spain7, representing high virulence isolates and Nc-Ger6, Nc-Spain2H and Nc-Spain1H, representing low virulence isolates. Pairwise comparisons between all isolates and between high virulence and low virulence groups identified a subset of proteins with higher abundance in high virulence isolates. These proteins were involved in energy and redox metabolism, and DNA/RNA processing, which might determine the faster growth rates and parasite survival of the high virulence isolates. Highlighted proteins included a predicted member of the rhoptry kinase family ROP20 specific for N. caninum, Bradyzoite pseudokinase 1 and several dense granule proteins. DNA polymerase, which was more abundant in all high virulence isolates in all comparisons, might also be implicated in virulence. These results reveal insights into possible mechanisms involved in specific phenotypic traits and virulence in N. caninum, and the relevance of these candidate proteins for N. caninum virulence deserves further investigation.


Assuntos
Neospora/metabolismo , Proteoma/metabolismo , Proteínas de Protozoários/metabolismo , Virulência , Cromatografia Líquida , Metabolismo Energético , Estágios do Ciclo de Vida , Ácidos Nucleicos/metabolismo , Fenótipo , Proteômica/métodos , Espectrometria de Massas em Tandem
3.
Ginecol Obstet Mex ; 83(10): 614-26, 2015 Oct.
Artigo em Espanhol | MEDLINE | ID: mdl-26859923

RESUMO

Polycystic ovary syndrome (PCOS) is known as a common gynecologic and endocrinology disease with multiple short and long-term consequences. It is one of the most common causes for hyperandrogenism and anovulation, increases the risk for metabolic syndrome, type 2 diabetes and cardiovascular disease. Its etiology remains unclear. PCOS is thought to be the result of the interaction between predisposing genetic variants with environmental factors and strongly depends on ethnicity. Proteomics allows the study of several hundreds or thousands of proteins in order to reveal physiological state of a tissue or an organ at the molecular level and to identify disease-specific biomarkers. Its use on PCOS patients will permit us to identify molecules that are involved in the PCOS pathology so we can develop specific diagnostic and management approaches.


Assuntos
Biomarcadores/metabolismo , Síndrome do Ovário Policístico/fisiopatologia , Proteômica/métodos , Feminino , Predisposição Genética para Doença , Humanos , Síndrome do Ovário Policístico/complicações , Síndrome do Ovário Policístico/etiologia
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