72 research outputs found
Genotypic Characterization of Streptococcus canis Isolated from Distinct Hosts with Special Emphasis on Multilocus Sequence Typing
Background: The animal pathogen Streptococcus canis is increasingly being noticed in human infections. Our aim was to develop a new multilocus sequence typing (MLST) scheme for Streptococcus canis and to compare isolates recovered from house pets and humans, in order to define the clonal structure of the S. canis population and explore the zoonotic potential of distinct S. canis genetic lineages.
Methods: Eighty-five S. canis isolates recovered from infections in animals (n = 78, recovered from 2000 to 2010 in three European countries, mainly from house pets) and humans (n = 7, recovered from 2006 to 2010 in Portugal) were studied. Isolates were identified by API 20 Strep, 23S rRNA gene targeted PCR and 16S rRNA gene sequencing, and characterized by MLST, pulsed-field gel electrophoresis (PFGE) and emm typing.
Results: All isolates were successfully typed with the proposed MLST scheme, indicating its applicability to S. canis from distinct sources. The MLST analysis showed a polyclonal structure of the S. canis population, where the same genetic lineages are found infecting house pets and humans and are disseminated in distinct geographic locations. PFGE confirmed the MLST findings, as it identified the same prevailing lineages and further strengthened the similarity between animal and human isolates. Phylogenetic analysis conducted with the 16S rRNA and MLST loci sequence data indicated that S. canis was a divergent taxon of the sister species Streptococcus pyogenes and Streptococcus dysgalactiae subsp. equisimilis, and found evidence of acquisition of genetic material by S. canis from the latter species. The presence of emm-like genes was restricted to a few isolates and correlated with MLST defined genetic lineages.
Conclusion: Our data shows that S. canis isolated from house pets and humans are a single population, and demonstrates that isolates belonging to the main genetic lineages identified are able to infect the human host, providing strong evidence for the zoonotic nature of S. canis infection in humans. A MLST database for S. canis was established at http://pubmlst.org/scanis/ (hosted by the Department of Zoology, University of Oxford, United Kingdom), constituting a valuable tool for future studies on the molecular epidemiology of this pathogen
Beta-hemolytic Streptococcus dysgalactiae strains isolated from horses are a genetically distinct population within the Streptococcus dysgalactiae taxon
The pathogenic role of beta-hemolytic Streptococcus dysgalactiae in the equine
host is increasingly recognized. A collection of 108 Lancefield group C (n =
96) or L (n = 12) horse isolates recovered in the United States and in three
European countries presented multilocus sequence typing (MLST) alleles,
sequence types and emm types (only 56% of the isolates could be emm typed)
that were, with few exceptions, distinct from those previously found in human
Streptococcus dysgalactiae subsp. equisimilis. Characterization of a subset of
horse isolates by multilocus sequence analysis (MLSA) and 16S rRNA gene
sequence showed that most equine isolates could also be differentiated from S.
dysgalactiae strains from other animal species, supporting the existence of a
horse specific genomovar. Draft genome information confirms the
distinctiveness of the horse genomovar and indicates the presence of
potentially horse-specific virulence factors. While this genomovar represents
most of the isolates recovered from horses, a smaller MLST and MLSA defined
sub-population seems to be able to cause infections in horses, other animals
and humans, indicating that transmission between hosts of strains belonging to
this group may occur
Beta-hemolytic Streptococcus dysgalactiae strains isolated from horses are a genetically distinct population within the Streptococcus dysgalactiae taxon
The pathogenic role of beta-hemolytic Streptococcus dysgalactiae in the equine host is increasingly
recognized. A collection of 108 Lancefield group C (n=96) or L (n=12) horse isolates recovered in
the United States and in three European countries presented multilocus sequence typing (MLST)
alleles, sequence types and emm types (only 56% of the isolates could be emm typed) that were, with
few exceptions, distinct from those previously found in human Streptococcus dysgalactiae subsp.
equisimilis. Characterization of a subset of horse isolates by multilocus sequence analysis (MLSA) and
16S rRNA gene sequence showed that most equine isolates could also be differentiated from
S. dysgalactiae strains from other animal species, supporting the existence of a horse specific
genomovar. Draft genome information confirms the distinctiveness of the horse genomovar and
indicates the presence of potentially horse-specific virulence factors. While this genomovar represents
most of the isolates recovered from horses, a smaller MLST and MLSA defined sub-population seems to
be able to cause infections in horses, other animals and humans, indicating that transmission between
hosts of strains belonging to this group may occur
Streptococcus canis Are a Single Population Infecting Multiple Animal Hosts Despite the Diversity of the Universally Present M-Like Protein SCM
Streptococcus canis is an animal pathogen which occasionally causes infections in humans. The S. canis M-like protein (SCM) encoded by the scm gene, is its best characterized virulence factor but previous studies suggested it could be absent in a substantial fraction of isolates. We studied the distribution and variability of the scm gene in 188 S. canis isolates recovered from companion animals (n = 152), wild animal species (n = 20), and humans (n = 14). Multilocus sequence typing, including the first characterization of wildlife isolates, showed that the same lineages are present in all animal hosts, raising the possibility of extensive circulation between species. Whole-genome analysis revealed that emm-like genes found previously in S. canis correspond to divergent scm genes, indicating that what was previously believed to correspond to two genes is in fact the same scm locus. We designed primers allowing for the first time the successful amplification of the scm gene in all isolates. Analysis of the scm sequences identified 12 distinct types, which could be divided into two clusters: group I (76%, n = 142) and group II (24%, n = 46) sharing little sequence similarity. The predicted group I SCM showed extensive similarity with each other outside of the N-terminal hypervariable region and a conserved IgG binding domain. This domain was absent from group II SCM variants found in isolates previously thought to lack the scm gene, which also showed greater amino acid variability. Further studies are necessary to elucidate the possible host interacting partners of the group II SCM variants and their role in virulence.Additional co-authors listed: Portuguese Group for the Study of Streptococcal Infection
Population Genetics of Streptococcus dysgalactiae Subspecies equisimilis Reveals Widely Dispersed Clones and Extensive Recombination
Streptococcus dysgalactiae subspecies equisimilis (SDSE) is an emerging global pathogen that can colonize and infect humans. Although most SDSE isolates possess the Lancefield group G carbohydrate, a significant minority have the group C carbohydrate. Isolates are further sub-typed on the basis of differences within the emm gene. To gain a better understanding of their molecular epidemiology and evolutionary relationships, multilocus sequence typing (MLST) analysis was performed on SDSE isolates collected from Australia, Europe and North America.The 178 SDSE isolates, representing 37 emm types, segregate into 80 distinct sequence types (STs) that form 17 clonal complexes (CCs). Eight STs recovered from all three continents account for >50% of the isolates. Thus, a small number of STs are highly prevalent and have a wide geographic distribution. Both ST and CC strongly correlate with group carbohydrate. In contrast, eleven STs were associated with >1 emm type, suggestive of recombinational replacements involving the emm gene; furthermore, 35% of the emm types are associated with genetically distant STs. Data also reveal a history of extensive inter- and intra-species recombination involving the housekeeping genes used for MLST. Sequence analysis of single locus variants identified through goeBURST indicates that genetic change mediated by recombination occurred approximately 4.4 times more frequently than by point mutation.A few genetic lineages with an intercontinental distribution dominate among SDSE causing infections in humans. The distinction between group C and G isolates reflects recent evolution, and no long-term genetic isolation between them was found. Lateral gene transfer and recombination involving housekeeping genes and the emm gene are important mechanisms driving genetic variability in the SDSE population
Prevention of hypertension in patients with pre-hypertension: protocol for the PREVER-prevention trial
<p>Abstract</p> <p>Background</p> <p>Blood pressure (BP) within pre-hypertensive levels confers higher cardiovascular risk and is an intermediate stage for full hypertension, which develops in an annual rate of 7 out of 100 individuals with 40 to 50 years of age. Non-drug interventions to prevent hypertension have had low effectiveness. In individuals with previous cardiovascular disease or diabetes, the use of BP-lowering agents reduces the incidence of major cardiovascular events. In the absence of higher baseline risk, the use of BP agents reduces the incidence of hypertension. The PREVER-prevention trial aims to investigate the efficacy, safety and feasibility of a population-based intervention to prevent the incidence of hypertension and the development of target-organ damage.</p> <p>Methods</p> <p>This is a randomized, double-blind, placebo-controlled clinical trial, with participants aged 30 to 70 years, with pre-hypertension. The trial arms will be chlorthalidone 12.5 mg plus amiloride 2.5 mg or identical placebo. The primary outcomes will be the incidence of hypertension, adverse events and development or worsening of microalbuminuria and of left ventricular hypertrophy in the EKG. The secondary outcomes will be fatal or non-fatal cardiovascular events: myocardial infarction, stroke, heart failure, evidence of new sub-clinical atherosclerosis, and sudden death. The study will last 18 months. The sample size was calculated on the basis of an incidence of hypertension of 14% in the control group, a size effect of 40%, power of 85% and P alpha of 5%, resulting in 625 participants per group. The project was approved by the Ethics committee of each participating institution.</p> <p>Discussion</p> <p>The early use of blood pressure-lowering drugs, particularly diuretics, which act on the main mechanism of blood pressure rising with age, may prevent cardiovascular events and the incidence of hypertension in individuals with hypertension. If this intervention shows to be effective and safe in a population-based perspective, it could be the basis for an innovative public health program to prevent hypertension in Brazil.</p> <p>Trial Registration</p> <p>Clinical Trials <a href="http://www.clinicaltrials.gov/ct2/show/NCT00970931">NCT00970931</a>.</p
Enhancement of metastatic ability by ectopic expression of ST6GalNAcI on a gastric cancer cell line in a mouse model
ST6GalNAcI is a sialyltransferase responsible for the synthesis of sialyl Tn (sTn) antigen which is expressed in a variety of adenocarcinomas including gastric cancer especially in advanced cases, but the roles of ST6GalNAcI and sTn in cancer progression are largely unknown. We generated sTn-expressing human gastric cancer cells by ectopic expression of ST6GalNAcI to evaluate metastatic ability of these cells and prognostic effect of ST6GalNAcI and sTn in a mouse model, and identified sTn carrier proteins to gain insight into the function of ST6GalNAcI and sTn in gastric cancer progression. A green fluorescent protein-tagged human gastric cancer cell line was transfected with ST6GalNAcI to produce sTn-expressing cells, which were transplanted into nude mice. STn-positive gastric cancer cells showed higher intraperitoneal metastatic ability in comparison with sTn-negative control, resulting in shortened survival time of the mice, which was mitigated by anti-sTn antibody administration. Then, sTn-carrying proteins were immunoprecipitated from culture supernatants and lysates of these cells, and identified MUC1 and CD44 as major sTn carriers. It was confirmed that MUC1 carries sTn also in human advanced gastric cancer tissues. Identification of sTn carrier proteins will help understand mechanisms of metastatic phenotype acquisition of gastric cancer cells by ST6GalNAcI and sTn
CU$TO CAPRINO CORTE 1.0: software de controle de custos para a caprinocultura de corte
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