BackgroundA large collection of sequenced mycobacteriophages capable of infecting a single host strain of Mycobacterium smegmatis shows considerable genomic diversity with dozens of distinctive types (clusters) and extensive variation within those sharing evident nucleotide sequence similarity. Here we profiled the mycobacterial components of a large composting system at the São Paulo zoo.ResultsWe isolated and sequenced eight mycobacteriophages using Mycobacterium smegmatis mc2155 as a host. None of these eight phages infected any of mycobacterial strains isolated from the same materials. The phage isolates span considerable genomic diversity, including two phages (Barriga, Nhonho) related to Subcluster A1 phages, two Cluster B phages (Pops, Subcluster B1; Godines, Subcluster B2), three Subcluster F1 phages (Florinda, Girafales, and Quico), and Madruga, a relative of phage Patience with which it constitutes the new Cluster U. Interestingly, the two Subcluster A1 phages and the three Subcluster F1 phages have genomic relationships indicating relatively recent evolution within a geographically isolated niche in the composting system.ConclusionsWe predict that composting systems such as those used to obtain these mycobacteriophages will be a rich source for the isolation of additional phages that will expand our view of bacteriophage diversity and evolution.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-016-0734-3) contains supplementary material, which is available to authorized users.
hOutbreaks of infections by rapidly growing mycobacteria following invasive procedures, such as ophthalmological, laparoscopic, arthroscopic, plastic, and cardiac surgeries, mesotherapy, and vaccination, have been detected in Brazil since 1998. Members of the Mycobacterium chelonae-Mycobacterium abscessus group have caused most of these outbreaks. As part of an epidemiological investigation, the isolates were typed by pulsed-field gel electrophoresis (PFGE). In this project, we performed a large-scale comparison of PFGE profiles with the results of a recently developed multilocus sequence typing (MLST) scheme for M. abscessus. Ninety-three isolates were analyzed, with 40 M. abscessus subsp. abscessus isolates, 47 M. abscessus subsp. bolletii isolates, and six isolates with no assigned subspecies. Forty-five isolates were obtained during five outbreaks, and 48 were sporadic isolates that were not associated with outbreaks. For MLST, seven housekeeping genes (argH, cya, glpK, gnd, murC, pta, and purH) were sequenced, and each isolate was assigned a sequence type (ST) from the combination of obtained alleles. The PFGE patterns of DraI-digested DNA were compared with the MLST results. All isolates were analyzable by both methods. Isolates from monoclonal outbreaks showed unique STs and indistinguishable or very similar PFGE patterns. Thirty-three STs and 49 unique PFGE patterns were identified among the 93 isolates. The Simpson's index of diversity values for MLST and PFGE were 0.69 and 0.93, respectively, for M. abscessus subsp. abscessus and 0.96 and 0.97, respectively, for M. abscessus subsp. bolletii. In conclusion, the MLST scheme showed 100% typeability and grouped monoclonal outbreak isolates in agreement with PFGE, but it was less discriminative than PFGE for M. abscessus.
Anti-idiotype antibodies have been considered for vaccination approaches against different diseases, including cancers. Based on that, we previously described an anti-bevacizumab idiotype monoclonal antibody, 10.D7, that revealed detectable antitumor effects on a vascular endothelial growth factor (VEGF)-dependent tumor model. Herein, we evaluated the possible applicability of a single-chain variable fragment (scFv) for the 10.D7 antibody in a gene immunization strategy. After checking that mammalian cells transfected to express the 10.D7 scFv are recognized by bevacizumab, it was explored the ability of our scFv construction, in a gene-based scheme, to elicit an immune response containing VEGF-binding antibodies. The results provide evidence that the designed 10.D7 scFv construct maintains the anti-bevacizumab idiotype features and has potential to activate an immune response recognizing VEGF.
r e v b r a s o r t o p . 2 0 1 4;4 9(6):642-646 w w w . r b o . o r g . b r Artigo Original Perfil de expressão de genes do colágeno na cápsula glenoumeral de pacientes com instabilidade traumática anterior do ombro ଝ informações sobre o artigo Histórico do artigo: Recebido em 22 de julho de 2013 Aceito em 21 de outubro de 2013 On-line em 26 de junho de 2014 Palavras-chave: Instabilidade do ombro Cápsula articular Expressão gênica Matriz extracelular Colágeno r e s u m o Objetivo: Avaliar a expressão dos genes COL1A1, COL1A2, COL3A1 e COL5A1 na cápsula glenoumeral de pacientes com instabilidade anterior traumática do ombro. Métodos: Foram avaliadas amostras de cápsula glenoumeral de 18 pacientes com instabilidade anterior traumática do ombro. Foram incluídos pacientes masculinos, com teste de apreensão positivo e lesão de Bankart no exame de ressonância magnética. Todos os pacientes sofreram mais de um episódio de luxação do ombro. Foram coletadas amostras da cápsula glenoumeral lesionada (região anteroinferior) e da região macroscopicamente não afetada (região anterossuperior) de cada paciente. A expressão dos genes de colágeno foi avaliada por reação em cadeia da polimerase após transcrição reversa com análise quantitativa (qRT-PCR).Resultados: A expressão de COL1A1, COL1A2 e COL3A1 não diferiu entre as duas regiões da cápsula do ombro. No entanto, foi observado que a expressão de COL5A1 estava significantemente reduzida na região anteroinferior em relação à região anterossuperior (mediana ± intervalo interquartílico: 0,057 ± 0,052 vs 0,155 ± 0,398; p = 0,028) da cápsula glenoumeral.Conclusão: A região afetada da cápsula glenoumeral de pacientes com instabilidade do ombro apresentou uma expressão reduzida de COL5A1. a b s t r a c tObjective: To evaluate the expression of the genes COL1A1, COL1A2, COL3A1 and COL5A1 in the glenohumeral capsule of patients with traumatic anterior instability of the shoulder.
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