RESUMEN
Candida inconspicua and Candida (Pichia) norvegensis are two emerging pathogenic species that exhibit reduced susceptibility to azole derivatives. Conventional (biochemical) approaches do not readily differentiate between the two species. The first aim of this work was to analyze the performance of biochemical, proteomic (matrix-assisted laser desorption ionization-time of flight [MALDI-TOF]), and molecular approaches in the precise identification of these species. These results then led us to sequence 3 genomic loci, i.e., the internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA), the D1/D2 domain of the 28S rDNA, and the elongation factor 1α (EF-1α) gene, either directly or following cloning, of 13 clinical isolates and 9 reference strains belonging to the 5 species included in the Pichia cactophila clade, namely, Pichia cactophila, Pichia insulana, C. inconspicua, C. norvegensis, and P. pseudocactophila. Finally, isolates of C. inconspicua were challenged for sexual reproduction on the appropriate medium. Our results show that EF-1α sequencing and proteic profiling by MALDI-TOF are the two most efficient approaches to distinguish between C. norvegensis and C. inconspicua. As a characteristic of the P. cactophila clade, we found multiple alleles of the rDNA regions in certain strains belonging to the tested species, making ITS or D1/D2 sequencing not appropriate for identification. Whatever the method of identification, including MALDI-TOF and EF-1α sequencing, none could differentiate C. inconspicua from P. cactophila. The results of phylogenetic analysis and the generation of asci from pure cultures of all C. inconspicua strains both support the identification of P. cactophila as the teleomorph of C. inconspicua.
Asunto(s)
Candida/clasificación , Cruzamientos Genéticos , Orden Génico , Candida/química , Candida/genética , Candida/metabolismo , Candidiasis/microbiología , Análisis por Conglomerados , ADN de Hongos/química , ADN de Hongos/genética , ADN Ribosómico/química , ADN Ribosómico/genética , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Humanos , Datos de Secuencia Molecular , Técnicas de Tipificación Micológica , Factor 1 de Elongación Peptídica/genética , Filogenia , Proteoma/análisis , ARN Ribosómico 28S/genética , Análisis de Secuencia de ADN , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Decompressive craniectomy has been revived as a radical surgical treatment in increased ICP. While the life saving effect is undisputable, reports about the functional outcome are controversial. Furthermore, there are no data comparing the outcome of craniectomised patients with different aetiologies. In a retrospective and prospective study we assessed the functional outcome of craniectomised patients (41 stroke and 24 traumatic brain injury (TBI) patients). The majority of patients considerably benefits from the rehabilitation process. Nevertheless, most of them remained dependent upon caregivers. In stroke patients there was a significant negative correlation between age and outcome. Even though TBI were younger than stroke patients there were no outcome differences between these groups. We found no correlation between the side of the hemisphere injured and outcome.