Intraclonal Genome Stability of the Metallo-beta-lactamase SPM-1-producing Pseudomonas aeruginosa ST277, an Endemic Clone Disseminated in Brazilian Hospitals

Carbapenems represent the mainstay therapy for the treatment of serious P aeruginosa infections. However, the emergence of carbapenem resistance has jeopardized the clinical use of this important class of compounds. The production of SPM-1 metallo-beta-lactamase has been the most common mechanism of carbapenem resistance identified in P. aeruginosa isolated from Brazilian medical centers. Interestingly, a single SPM-1-producing P. aeruginosa clone belonging to the ST277 has been widely spread within the Brazilian territory. In the current study, we performed a next-generation sequencing of six SPM-1-producing P. aeruginosa ST277 isolates. The core genome contains 5899 coding genes relative to the reference strain P. aeruginosa PAO1. A total of 26 genomic islands were detected in these isolates. We identified remarkable elements inside these genomic islands, such as copies of the bla(spM-1) gene conferring resistance to carbapenems and a type I-C CRISPR-Cas system, which is involved in protection of the chromosome against foreign DNA. In addition, we identified single nucleotide polymorphisms causing amino acid changes in antimicrobial resistance and virulence-related genes. Together,these factors could contribute to the marked resistance and persistence of the SPM-1-producing P aeruginosa ST277 clone. A comparison of the SPM-1-producing P. aeruginosa ST277 genomes showed that their core genome has a high level nucleotide similarity and synteny conservation. The variability observed was mainly due to acquisition of genomic islands carrying several antibiotic resistance genes.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Lab Nacl Comp Cient, Lab Bioinformat, Petropolis, BrazilUniv Fed Sao Paulo, Escola Paulista Med, Dept Internal Med, Lab Alerta,Div Infect Dis, Sao Paulo, BrazilLaboratório Alerta, Division of Infectious Diseases, Department of Internal Medicine, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, BrazilCNPq: 305535/2014-5CNPq: 302768/2011-4CNPq: 312864/2015-9FAPERJ: E-26/202.903/2016Web of Scienc

Exploring the long-term associations between adolescents’ music training and academic achievement

There is a positive relationship between learning music and academic achievement, although doubts remain regarding the mechanisms underlying this association. This research analyses the academic performance of music and non-music students from seventh to ninth grade. The study controls for socioeconomic status, intelligence, motivation and prior academic achievement. Data were collected from 110 adolescents at two time points, once when the students were between 11 and 14 years old in the seventh grade, and again 3 years later. Our results show that music students perform better academically than non-music students in the seventh grade (Cohen’s d = 0.88) and in the ninth grade (Cohen’s d = 1.05). This difference is particularly evident in their scores in Portuguese language and natural science; the difference is somewhat weaker in history and geography scores, and is least pronounced in mathematics and English scores (η2p from .09 to .21). A longitudinal analysis also revealed better academic performance by music students after controlling for prior academic achievement (η2p = .07). Furthermore, controlling for intelligence, socioeconomic status and motivation did not eliminate the positive association between music learning from the seventh to the ninth grade and students’ academic achievement (η2p = .06). During the period, music students maintained better and more consistent academic standing. We conclude that, after controlling for intelligence, socioeconomic status and motivation, music training is positively associated with academic achievement.This research was funded by the Portuguese National Funding Agency for Science, Research and Technology (FCT - Fundacao para a Ciencia e a Tecnologia)

Clinical and molecular description of a high-copy IncQ1 KPC-2 plasmid harbored by the international ST15 Klebsiella pneumoniae clone

This study provides the genomic characterization and clinical description of bloodstream infections (BSI) cases due to ST15 KPC-2 producer Klebsiella pneumoniae. Six KPC-K. pneumoniae isolates were recovered in 2015 in a tertiary Brazilian hospital and were analyzed by whole-genome sequencing (WGS) (Illumina MiSeq short reads). Of these, two isolates were further analyzed by Nanopore MinION sequencing, allowing complete chromosome and plasmid circularization (hybrid assembly), using Unicycler software. The clinical analysis showed that the 30-day overall mortality for these BSI cases was high (83%). The isolates exhibited meropenem resistance (MICs, 32 to 128 mg/liter), with 3/6 isolates resistant to polymyxin B. The conjugative properties of the blaKPC-2 plasmid and its copy number were assessed by standard conjugation experiments and sequence copy number analysis. We identified in all six isolates a small (8.3-kb), high-copy-number (20 copies/cell) non-self-conjugative IncQ plasmid harboring blaKPC-2 in a non-Tn4401 transposon. This plasmid backbone was previously reported to harbor blaKPC-2 only in Brazil, and it could be comobilized at a high frequency (10−4) into Escherichia coli J53 and into several high-risk K. pneumoniae clones (ST258, ST15, and ST101) by a common IncL/M helper plasmid, suggesting the potential of international spread. This study thus identified the international K. pneumoniae ST15 clone as a carrier of blaKPC-2 in a high-copy-number IncQ1 plasmid that is easily transmissible among other common Klebsiella strains. This finding is of concern since IncQ1 plasmids are efficient antimicrobial resistance determinant carriers across Gram-negative species. The spread of such carbapenemase-encoding IncQ1 plasmids should therefore be closely monitored. IMPORTANCE In many parts of the world, carbapenem resistance is a serious public health concern. In Brazil, carbapenem resistance in Enterobacterales is mostly driven by the dissemination of KPC-2-producing K. pneumoniae clones. Despite being endemic in this country, only a few reports providing both clinical and genomic data are available in Brazil, which limit the understanding of the real clinical impact caused by the dissemination of different clones carrying blaKPC-2 in Brazilian hospitals. Although several of these KPC-2-producer K. pneumoniae isolates belong to the clonal complex 258 and carry Tn4401 transposons located on large plasmids, a concomitant emergence and silent dissemination of small high-copy-number blaKPC-2 plasmids are of importance, as described in this study. Our data identify a small high-copy-number IncQ1 KPC plasmid, its clinical relevance, and the potential for conjugative transfer into several K. pneumoniae isolates, belonging to different international lineages, such as ST258, ST101, and ST15

Bologna process, higher education and a few considerations about the New University

O presente artigo analisa o que se convencionou chamar de Processo de Bolonha, isto é, a produção de uma “política pública de um meta-Estado para um meta-campo universitário”, constituindo-se em uma política educacional supranacional, comum aos estados-membros da União Européia, com vista à construção de um “espaço europeu de educação superior”. O processo político e de reformas institucionais, realizado por cada governo nacional, conduzirá ao estabelecimento efetivo do novo sistema europeu de educação superior até 2010, incluindo atualmente 45 países – todos os da UE e outros 18 países europeus não pertencentes a ela. Nesse sentido, por se tratar de um vastíssimo número de “subsistemas nacionais” e de instituições educativas, atribui-se um grande protagonismo às questões relativas à “garantia de qualidade”. Analisam-se, igualmente, as recentes transformações na educação superior no Brasil, em que o projeto da chamada “Universidade Nova” e o Programa de Apoio a Planos de Reestruturação e Expansão das Universidades Federais (REUNI) constituem-se nas manifestações mais claras do reordenamento desse nível de ensino (seguindo os parâmetros de Bolonha), que já experimentara grandes transformações nos governos de Fernando Henrique Cardoso (1995-2002) e teve prosseguimento nos governos de Luiz Inácio Lula da Silva (2003-2006; 2007), embora com distintos matizes.This article analyzes what is conventionally known as the Bologna Process, or the making of a “public policy of a meta-State for a University meta-field” that corresponds to a supranational educational policy for all the European Union membership States, with the goal of building a “European higher education space.” The political process and the institutional reforms of each national government intends to establish the new European higher education system until 2010, with 45 countries – the number reflects current developments, including the EU membership States and 18 non-EU countries. Given the high quantity and the myriads of “national subsystems” and educational institutions involved, “quality assurance” becomes a major task in this process. We analyze, in the same way, the recent higher education changes in Brazil, where the so-called “New University” project and the Program of Support for the Restructuring and Expansion of Brazilian Federal Universities (REUNI, in Portuguese) are the clearest expressions of the reshaping of the higher education system (in accordance with the Bologna standards) after the dramatic changes made by Fernando Henrique Cardoso´s government (1995-2002) and continued by Luiz Inácio Lula da Silva´s government (2003-2006; 2007), despite some differences between both administrations

Relative expression of mRNAs related to cavitation process in bovine embryos produced in vivo and in vitro

The objectives of this work were to identify and to evaluate possible differences on gene expression of aquaporins and Na/K-ATPases transcripts between embryos in vivo and in vitro produced. For each group, 15 blastocysts distributed in three pools were used for RNA extraction followed by amplification and reverse transcription. The resulting cDNAs were submitted to Real-Time PCR, using the GAPDH gene as endogenous control. It was not possible to identify AQP1 transcripts. Relative expression of AQP3 (1.33 ± 0.78) and AQP11 (2.00 ± 1.42) were not different in blastocysts in vitro and in vivo produced. Na/K-ATPase α1 gene (2.25 ± 1.07) was overregulated whereas Na/K-ATPase β2 transcripts 0.40 ± 0.30) did not differ among blastocysts produced in vitro from those produced in vivo. Transcripts for gene AQP1 are not present in bovine blastocysts. In vitro culture system does not alter expression of genes AQP3, AQP11 and Na/K-ATPase β2 genes, however, it affects expression of Na/K-ATPase α1.Os objetivos neste trabalho foram identificar e avaliar possíveis diferenças na expressão gênica de transcritos de Aquaporina e ATPases-Na/K presentes em embriões produzidos in vivo e in vitro. Para cada grupo, 15 blastocistos distribuídos em três conjuntos foram utilizados para a extração do RNA, seguida da amplificação e da transcrição reversa. Os DNAs complementares foram submetidos à reação em cadeia da enzima polimerase em tempo real, utilizando-se o gene GAPDH como controle endógeno. Não foi possível identificar transcritos de AQP1. A expressão relativa dos genes AQP3 (1,33 ± 0,78) e AQP11 (2,00 ± 1,42) não foi diferente em blastocistos produzidos in vitro e in vivo. O gene ATPase-Na/K α1 (2,25 ± 1,07) encontrou-se sobrerregulado, enquanto o gene ATPase-Na/K β2 (0,40 ± 0,30) não diferiu entre os blastocistos produzidos in vitro e aqueles produzidos in vivo. Transcritos para o gene AQP1 não estão presentes em blastocistos bovinos. O sistema de cultivo in vitro não influencia a expressão dos genes AQP3, AQP11 e ATPase-Na/K β2, porém altera a expressão do gene ATPase-Na/K α1

Optical phonon features of triclinic montebrasite : dispersion analysis and non-polar Raman modes.

Polarized infrared and Raman spectra of triclinic LiAl(PO4)(OH) [montebrasite] single crystal were recorded for appropriate optical configurations. Dispersion analysis was applied on the infrared reflectivity spectra taken at low incidence angle (11 ) to determine the oscillator parameters and the dipole directions of the polar phonons. In particular, all the 27 polar phonons, predicted by group theory for triclinic P1 structure,were determined. The obtained dielectric tensor parameters have been checked by comparison between predicted and measured infrared spectra at higher incidence angle (34 ). The azimuth and co-elevation angles obtained from the dispersion analysis showed that the response of several polar phonons is close to that of an orthorhombic system. Polarized Raman spectra obtained in several scattering geometries allowed us to obtain well-defined 24 non-polar modes, also in perfect agreement with group theory. The selection rule between Raman and infrared phonons was respected, confirming the centrosymmetric structure and ruling out any relevant influence of defects. The relatively narrow phonon bands are compatible with a highly ordered structure with fully occupied atomic sites

Differential effects of antigens from L. braziliensis isolates from disseminated and cutaneous leishmaniasis on in vitro cytokine production

BACKGROUND: Disseminated leishmaniasis is an emerging infectious disease, mostly due to L. braziliensis, which has clinical and histopathological features distinct from cutaneous leishmaniasis. METHODS: In the current study we evaluated the in vitro production of the cytokines IFN-γ, TNF-α, IL-5 and IL-10 by peripheral blood mononuclear cells (PBMC) from 15 disseminated leishmaniasis and 24 cutaneous leishmaniasis patients upon stimulation with L. braziliensis antigens genotyped as disseminated leishmaniasis or cutaneous leishmaniasis isolates. RESULTS: Regardless of the source of L. braziliensis antigens, PBMC from cutaneous leishmaniasis patients produced significantly higher IFN-γ than PBMC from disseminated leishmaniasis patients. Levels of TNF-α by PBMC from cutaneous leishmaniasis patients were significantly higher than disseminated leishmaniasis patients only when stimulated by genotyped cutaneous leishmaniasis antigens. The levels of IL-5 and IL-10 production by PBMC were very low and similar in PBMCs from both disseminated leishmaniasis and cutaneous leishmaniasis patients. The immune response of each patient evaluated by the two L. braziliensis antigens was assessed in a paired analysis in which we showed that L. braziliensis genotyped as disseminated leishmaniasis isolate was more potent than L. braziliensis genotyped as cutaneous leishmaniasis isolate in triggering IFN-γ and TNF-α production in both diseases and IL-5 only in cutaneous leishmaniasis patients. CONCLUSION: This study provides evidence that antigens prepared from genotypically distinct strains of L. braziliensis induce different degrees of immune response. It also indicates that both parasite and host play a role in the outcome of L. braziliensis infection