Multiplex polymerase chain reaction to identify and determine the toxigenicity of Corynebacterium spp with zoonotic potential and an overview of human and animal infections

Corynebacterium diphtheriae, Corynebacterium ulcerans and Corynebacterium pseudotuberculosis constitute a group of potentially toxigenic microorganisms that are related to different infectious processes in animal and human hosts. Currently, there is a lack of information on the prevalence of disease caused by these pathogens, which is partially due to a reduction in the frequency of routine laboratory testing. In this study, a multiplex polymerase chain reaction (mPCR) assay that can simultaneously identify and determine the toxigenicity of these corynebacterial species with zoonotic potential was developed. This assay uses five primer pairs targeting the following genes: rpoB (Corynebacterium spp), 16S rRNA (C. ulcerans and C. pseudotuberculosis), pld (C. pseudotuberculosis), dtxR (C. diphtheriae) and tox [diphtheria toxin (DT) ]. In addition to describing this assay, we review the literature regarding the diseases caused by these pathogens. Of the 213 coryneform strains tested, the mPCR results for all toxigenic and non-toxigenic strains of C . diphtheriae, C. ulcerans and C. pseudotuberculosis were in 100% agreement with the results of standard biochemical tests and PCR-DT. As an alternative to conventional methods, due to its advantages of specificity and speed, the mPCR assay used in this study may successfully be applied for the diagnosis of human and/or animal diseases caused by potentially toxigenic corynebacterial species.

Study of Vaccinia and Cowpox viruses' replication in Rac1-N17 dominant-negative cells

Interfering with cellular signal transduction pathways is a common strategy used by many viruses to create a propitious intracellular environment for an efficient replication. Our group has been studying cellular signalling pathways activated by the orthopoxviruses Vaccinia (VACV) and Cowpox (CPXV) and their significance to viral replication. In the present study our aim was to investigate whether the GTPase Rac1 was an upstream signal that led to the activation of MEK/ERK1/2, JNK1/2 or Akt pathways upon VACV or CPXV' infections. Therefore, we generated stable murine fibroblasts exhibiting negative dominance to Rac1-N17 to evaluate viral growth and the phosphorylation status of ERK1/2, JNK1/2 and Akt. Our results demonstrated that VACV replication, but not CPXV, was affected in dominant-negative (DN) Rac1-N17 cell lines in which viral yield was reduced in about 10-fold. Viral late gene expression, but not early, was also reduced. Furthermore, our data showed that Akt phosphorylation was diminished upon VACV infection in DN Rac1-N17 cells, suggesting that Rac1 participates in the phosphoinositide-3 kinase pathway leading to the activation of Akt. In conclusion, our results indicate that while Rac1 indeed plays a role in VACV biology, perhaps another GTPase may be involved in CPXV replication.

Prevalence and genetic characterisation of HTLV-1 and 2 dual infections in patients with pulmonary tuberculosis in Central-West Brazil

Human T-cell lymphotropic virus (HTLV) may impact the clinical course of tuberculosis (TB). Both infections are highly endemic in Brazil. The aim of this study was to assess the prevalence of HTLV-1/2 in TB patients in Central-West Brazil and to perform a genetic characterisation of the respective isolates. Of the 402 patients, six (1.49%) were positive for anti-HTLV and five (1.24%; 95% confidence interval: 0.46-3.05) were infected with HTLV-1/2. Genetic characterisation demonstrated that the four HTLV-1 isolates belonged to the Transcontinental subgroup A of the Cosmopolitan subtype a and that the HTLV-2 isolate belonged to subtype a (HTLV-2a/c). The prevalence of HTLV infection observed in this study is higher than that observed in local blood donors and the HTLV-1 and 2 subtypes identified are consistent with those circulating in Brazil.

Oxidative stress enhances the expression of sulfur assimilation genes: preliminary insights on the Enterococcus faecalis iron-sulfur cluster machinery regulation

The Firmicutes bacteria participate extensively in virulence and pathological processes. Enterococcus faecalis is a commensal microorganism; however, it is also a pathogenic bacterium mainly associated with nosocomial infections in immunocompromised patients. Iron-sulfur [Fe-S] clusters are inorganic prosthetic groups involved in diverse biological processes, whose in vivo formation requires several specific protein machineries. Escherichia coli is one of the most frequently studied microorganisms regarding [Fe-S] cluster biogenesis and encodes the iron-sulfur cluster and sulfur assimilation systems. In Firmicutes species, a unique operon composed of the sufCDSUB genes is responsible for [Fe-S] cluster biogenesis. The aim of this study was to investigate the potential of the E. faecalis sufCDSUB system in the [Fe-S] cluster assembly using oxidative stress and iron depletion as adverse growth conditions. Quantitative real-time polymerase chain reaction demonstrated, for the first time, that Gram-positive bacteria possess an OxyR component responsive to oxidative stress conditions, as fully described for E. coli models. Likewise, strong expression of the sufCDSUB genes was observed in low concentrations of hydrogen peroxide, indicating that the lowest concentration of oxygen free radicals inside cells, known to be highly damaging to [Fe-S] clusters, is sufficient to trigger the transcriptional machinery for prompt replacement of [Fe-S] clusters.

Toxocara canis and the allergic process

The protective effect of infectious agents against allergic reactions has been thoroughly investigated. Current studies have demonstrated the ability of some helminths to modulate the immune response of infected hosts. The objective of the present study was to investigate the relationship between Toxocara canis infection and the development of an allergic response in mice immunised with ovalbumin (OVA). We determined the total and differential blood and bronchoalveolar lavage fluid cells using BALB/c mice as a model. To this end, the levels of interleukin (IL)-4, IL-5 and IL-10 and anti-OVA-IgE were measured using an ELISA. The inflammatory process in the lungs was observed using histology slides stained with haematoxylin and eosin. The results showed an increase in the total number of leukocytes and eosinophils in the blood of infected and immunised animals at 18 days after infection. We observed a slight lymphocytic inflammatory infiltrate in the portal space in all infected mice. Anti-OVA-IgE levels were detected in smaller proportions in the plasma of immunised and infected mice compared with mice that were only infected. Therefore, we concluded that T. canis potentiates inflammation in the lungs in response to OVA, although anti-OVA-IgE levels suggest a potential reduction of the inflammatory process through this mechanism.

Pesquisa em enfermagem e o positivismo

Discute-se os fundamentos do positivismo; as influências mais marcantes para a formulação da filosofia comtiana; como as idéias de Comte, tanto como doutrina, quanto como método, passam a dominar o pensamento da sociedade, ultrapassando o século XIX e alcançando o século XX; e como a doutrina e o método empírico positivista influenciaram a Enfermagem em sua noção acerca da ciência e de homem/ ambiente/doença.

Efeito da massagem prévia à punção venosa na reação do pré-escolar e escolar

Este trabalho tem por objetivo descrever o efeito da massagem efetuada pelos pais, na reação do pré-escolar e escolar hospitalizado, à punção venosa. Analisaram-se as reações das crianças quanto aos parâmetros de sinais vitais, verbalização e comunicação não-verbal. Os resultados obtidos mostraram que a massagem teve ação significativa nas reações não-verbais principalmente as relacionadas ao relaxamento muscular. Os efeitos sobre os sinais vitais não apresentaram alterações entre os dois procedimentos, sem massagem e com massagem, que foram realizados na mesma criança.

Qualidade de vida de estudantes de graduação em enfermagem

Este estudo teve como propósito investigar a qualidade de vida de 264 estudantes de Graduação em Enfermagem. Os dados foram coletados por meio de um questionário e um instrumento de medida de qualidade de vida, o IQV de Ferrans e Powers. As médias obtidas dos escores do IQV para os estudantes foram altas e próximas das médias da população geral de Rockford - USA. As médias do IQV nos diferentes anos do curso mostraram queda no decorrer do 2º ano, quando os estudantes são inseridos no campo prático, fornecendo subsídios para possíveis intervenções dos educadores nessa problemática.

O aprendizado da construção decaso clínico em saúde mental

Este estudo objetivou relatar a experiencia de uma aluna na construção de caso clinico em Saude Mental, desenvolvida nas disciplinas de Enfermagem em Saude Mental II e Enfermagem Psiquiátrica da Graduação em Enfermagem (Unicamp). Para estas disciplinas, a construção de caso é entendida como a articulação entre a entrevista psiquiátrica e alguns conceitos da teoria psicanalítica, aprendidos nas aulas, supervisao e leitura de textos, pretendendo estruturar um cuidado de Enfermagem coerente com a concepção de homem do referencial psicanalítico: um sujeito dividido pela existência do inconsciente, estruturado como linguagem. O relato baseou-se na experiência vivida pela aluna, durante as disciplinas, na produção de relatórios e construção de caso sob a forma de trabalho final. Foi possivel a aluna desenvolver a leitura do caso e construir uma teoria sustentadora do atendimento, por meio das demandas levantadas no diálogo com o paciente que direcionaram os pontos passíveis de intervenção.

O brincar em sala de espera de um Ambulatório Infantil: a visão dos profissionais de saúde

O objetivo deste estudo foi compreender, na perspectiva dos profissionais de saúde, o significado do uso do brincar/brinquedo em sala de espera de um ambulatório infantil. Realizaram-se entrevistas semiestruturadas com 11 trabalhadores. Procedeu-se à análise qualitativa dos dados, os quais foram organizados ao redor de dois temas: reconhecimento da importância do brincar/brinquedo e lidando com limitações. O brincar mostra-se como uma estratégia de cuidado à criança, pois ameniza o tempo de espera neste ambiente, altera positivamente o comportamento e valoriza o processo de desenvolvimento das mesmas, além de melhorar a comunicação e a interação com os profissionais da saúde. É preciso, entretanto, que exista uma estrutura física adequada do hospital, aliada aos profissionais de saúde e à equipe gestora sensíveis para a inclusão de novas estratégias de cuidado, a exemplo do brincar, a fim de humanizar a assistência à saúde da criança.

Assessment of the teaching-learning process in students of the health area: cardiopulmonary resuscitation maneuvers

OBJECTIVETo evaluate the skills and knowledge of undergraduate students in the health area on cardiopulmonary resuscitation maneuvers with the use of an automatic external defibrillator.METHODThe evaluation was performed in three different stages of the teaching-learning process. A theoretical and practical course was taught and the theoretical classes included demonstration. The evaluation was performed in three different stages of the teaching-learning process. Two instruments were applied to evaluate the skills (30-items checklist) and knowledge (40-questions written test). The sample comprised 84 students.RESULTSAfter the theoretical and practical course, an increase was observed in the number of correct answers in the 30-items checklist and 40-questions written test.CONCLUSIONAfter the theoretical class (including demonstration), only one of the 30-items checklist for skills achieved an index ≥ 90% of correct answers. On the other hand, an index of correct answers greater than 90% was achieved in 26 (86.7%) of the 30 items after a practical training simulation, evidencing the importance of this training in the defibrillation procedure.

Contrasting patterns of insecticide resistance and knockdown resistance (kdr) in Aedes aegypti populations from Jacarezinho (Brazil) after a Dengue Outbreak

ABSTRACT After a dengue outbreak, the knowledge on the extent, distribution and mechanisms of insecticide resistance is essential for successful insecticide-based dengue control interventions. Therefore, we evaluated the potential changes to insecticide resistance in natural Aedes aegypti populations to Organophosphates (OP) and Pyrethroids (PY) after chemical vector control interventions. After a Dengue outbreak in 2010, A. aegypti mosquitoes from the urban area of Jacarezinho (Paraná, Brazil) were collected in 2011 and 2012. Insecticide resistance to OP Temephos was assessed in 2011 and 2012 by dose–response bioassays adopting WHO-based protocols. Additionally, in both sampling, PY resistance was also investigated by the Val1016Ile mutation genotyping. In 2011, a random collection of mosquitoes was carried out; while in 2012, the urban area was divided into four regions where mosquitoes were sampled randomly. Bioassays conducted with larvae in 2011 (82 ± 10%; RR95 = 3.6) and 2012 (95 ± 3%; RR95 = 2.5) indicated an incipient altered susceptibility to Temephos. On the other hand, the Val1016IIe mutation analysis in 2011, presented frequencies of the 1016Ilekdr allele equal to 80%. Nevertheless, in 2012, when the urban area of Jacarezinho was analyzed as a single unit, the frequency of the mutant allele was 70%. Additionally, the distribution analysis of the Val1016Ile mutation in 2012 showed the mutant allele frequencies ≥60% in all regions. These outcomes indicated the necessity of developing alternative strategies such as insecticide rotations for delaying the evolution of resistance.