A new Trypanosoma cruzi genotyping method enables high resolution evolutionary analyses.

BACKGROUND: Trypanosoma cruzi is an important human pathogen in Latin America with nearly seven million people infected. It has a large degree of genetic diversity, classified into six discrete typing units (DTUs), which probably influences its physiological behavior and clinical manifestations. Several genotyping methods are available, with distinct performance on easiness, cost, resolution and applicability; no method excels in all parameters. OBJECTIVES AND METHODS: To devise a molecular method for T. cruzi genotyping, based on polymerase chain reaction (PCR) amplification of a single target with multiple copies in the nuclear genome by large scale sequencing. We have applied this method to 29 T. cruzi isolates, comprising all described DTUs. FINDINGS: We were able to classify all samples into sub DTU level with high robustness. Evolutionary relationship between DTUs were ascertained, suggesting that TcIII and TcIV DTUs are non-hybrid, and DTU IV is more similar to the common ancestral. CONCLUSION: As the TS-LSS method is based on a single PCR reaction, comprising several copies of the target, it is probably useful for clinical samples, when the amount of DNA is a limiting factor. As large scale sequencing systems become more common, the TS-LSS method can be increasingly applied for T. cruzi genotyping.

Exploring the parasite load and molecular diversity of Trypanosoma cruzi in patients with chronic Chagas disease from different regions of Brazil.

Chagas disease is still a major public health issue in many Latin American countries. One of the current major challenges is to find an association between Trypanosoma cruzi discrete typing units (DTUs) and clinical manifestations of the disease. In this study, we used a multilocus conventional PCR and quantitative real time PCR (qPCR) approaches to perform the molecular typing and parasite load quantification directly from blood specimens of 65 chronic Chagas disease patients. All patients were recruited at the same health center, but their place of birth were widely distributed in different geographic regions of Brazil. Of the 65 patients, 35 (53.8%) presented positive amplification by real time qPCR, being 20 (30.7%) with the clinical indeterminate form and 15 (23.1%) with the cardiac form of the disease. The parasite load median for all positive patients was 2.54 [1.43-11.14] parasite equivalents/mL (par. Eq./mL), with the load ranging from 0.12 to 153.66 par. Eq./mL. Noteworthy, the parasite load was significantly higher in patients over 70 years old (median 20.05 [18.29-86.86] par. Eq./mL). Using guanidine-EDTA blood samples spiked with reference T. cruzi strains, belonging to the six DTUs, it was possible to genotype the parasite up to 0.5 par. Eq./mL, with high specificity. Of the patients with positive qPCR, it was possible to identify the T. cruzi DTU in 28 patients (80%). For the remaining patients (20%), at least a partial result was obtained. Analysis of specimens showed prevalences of TcVI, TcII and mixed infection TcVI+TcII equal to 40%, 17.1% and 14.3%, respectively. In addition, two patients were infected by TcV, and one patient was coinfected by TcIII+TcVI, These last three patients were in stage A of chronic chagasic cardiomyopathy (CCC), and they were born at the Bahia State (northeast region of Brazil). When T. cruzi genotypes were compared with the parasite load, more elevated parasite loads were observed in patients infected by TcII in general (parasite load median of 7.56 par. Eq./mL) in comparison to patients infected by TcVI (median of 2.35 par. Eq./mL). However, while the frequency of CCC was 50% in patients infected by TcVI and TcV, only 16.7% of patients infected by TcII evolved to CCC. Taking together, our results contribute to update the epidemiological knowledge of T. cruzi DTUs in Brazil, and highlight the age of patient and infection by TcII as important features that lead to the observation of higher parasitemia levels.

[The use of virtual communities in the support to patients with breast cancer]. / Uso de comunidades virtuais no suporte a portadoras de câncer de mama.

Thinking about virtual communities as a space for interaction and the sharing of experiences, especially among individuals affected by chronic diseases, the scope of this study was to analyze the role and nature of support offered by virtual communities to breast cancer patients. It is a descriptive study with a qualitative approach. For data collection, the Netvizz application was used extracting data from the first five Facebook open breast cancer groups in the year 2016. There were 2921 posts, totaling 1722 comments. For data analysis a word cloud was created through WordArt, and the most recurrent words were discussed in cross-sectional categories: I, confessional, vs. you, interactional; therapeutic process, religiosity and the "Intervention Window." The results show that the support offered in these spaces occurs especially in the informational and emotional spheres, ranging from the diagnosis phase to the control of the disease. The understanding of the significance of the information shared in the virtual communities by women with breast cancer can contribute to the practice of care that is not restricted to the biological and technical approaches, but rather a more comprehensive and effective approach to holistic care.

Pensando nas comunidades virtuais como espaço de interação e compartilhamento de experiências, principalmente entre indivíduos acometidos por doenças crônicas, objetivamos com este estudo, analisar o papel e natureza de suporte oferecido pelas comunidades virtuais às portadoras de câncer de mama. Estudo descritivo com abordagem qualitativa. Para coleta de dados foi utilizado o aplicativo Netvizz, extraindo dados dos cinco primeiros grupos abertos de câncer de mama do Facebook, no ano de 2016. Foram encontradas 2921 postagens, totalizando 1722 comentários. Para análise dos dados foi criada uma nuvem de palavras através do WordArt, e as palavras mais recorrentes foram discutidas em categorias transversais: Eu Confessional X Você Interacional; processo terapêutico, religiosidade e "Arco de Intervenções". Constatamos, pelos resultados, que o suporte oferecido nestes espaços se dá especialmente nas esferas informacional e emocional, indo desde a fase de diagnóstico até o controle da doença. A compreensão das significações das informações partilhadas nas comunidades virtuais por mulheres com câncer de mama pode contribuir para a realização de práticas de cuidado que não se restrinjam às abordagens de cunho biológico e tecnicista, mas sim numa assistência holística, integral e efetiva.

Uso de comunidades virtuais no suporte a portadoras de câncer de mama / The use of virtual communities in the support to patients with breast cancer

Resumo Pensando nas comunidades virtuais como espaço de interação e compartilhamento de experiências, principalmente entre indivíduos acometidos por doenças crônicas, objetivamos com este estudo, analisar o papel e natureza de suporte oferecido pelas comunidades virtuais às portadoras de câncer de mama. Estudo descritivo com abordagem qualitativa. Para coleta de dados foi utilizado o aplicativo Netvizz, extraindo dados dos cinco primeiros grupos abertos de câncer de mama do Facebook, no ano de 2016. Foram encontradas 2921 postagens, totalizando 1722 comentários. Para análise dos dados foi criada uma nuvem de palavras através do WordArt, e as palavras mais recorrentes foram discutidas em categorias transversais: Eu Confessional X Você Interacional; processo terapêutico, religiosidade e "Arco de Intervenções". Constatamos, pelos resultados, que o suporte oferecido nestes espaços se dá especialmente nas esferas informacional e emocional, indo desde a fase de diagnóstico até o controle da doença. A compreensão das significações das informações partilhadas nas comunidades virtuais por mulheres com câncer de mama pode contribuir para a realização de práticas de cuidado que não se restrinjam às abordagens de cunho biológico e tecnicista, mas sim numa assistência holística, integral e efetiva.

Abstract Thinking about virtual communities as a space for interaction and the sharing of experiences, especially among individuals affected by chronic diseases, the scope of this study was to analyze the role and nature of support offered by virtual communities to breast cancer patients. It is a descriptive study with a qualitative approach. For data collection, the Netvizz application was used extracting data from the first five Facebook open breast cancer groups in the year 2016. There were 2921 posts, totaling 1722 comments. For data analysis a word cloud was created through WordArt, and the most recurrent words were discussed in cross-sectional categories: I, confessional, vs. you, interactional; therapeutic process, religiosity and the "Intervention Window." The results show that the support offered in these spaces occurs especially in the informational and emotional spheres, ranging from the diagnosis phase to the control of the disease. The understanding of the significance of the information shared in the virtual communities by women with breast cancer can contribute to the practice of care that is not restricted to the biological and technical approaches, but rather a more comprehensive and effective approach to holistic care.

Trichophoromyia auraensis is a putative vector

The sandfly Trichophoromyia auraensis has recently evolved as a proven vector of Leishmania (Viannia) endemic to state of Acre in the north of Brazil. This note is intended to propose a correction in the report of the first occurrence of natural infection of Leishmania (Viannia) in this species. We and the other scientific groups reinforced that Tr. auraensis is a possible vector involved in the transmission of American cutaneous leishmaniasis in Acre, Brazil.

Multiplex PCR as a tool for the diagnosis of Leishmania spp. kDNA and the gapdh housekeeping gene of mammal hosts.

BACKGROUND: The PCR assays usually employed for Leishmania diagnosis does not simultaneously detect a constitutive gene that would certify the viability of the DNA sample. We present a multiplex PCR approach for the simultaneous diagnosis of the Leishmania sp. kDNA fragment and a catalytic domain segment of a conserved region of the mammalian gapdh gene. METHODOLOGY: The proposed multiplex protocol was designed through in silico PCR. The annealing temperature, concentration of primer pairs, number of cycles, distinct polymerase enzymes and premix kit were defined to achieve an optimal reaction. The DNA detection sensitivity was tested with different concentrations of known L. tropica DNA, and the reproducibility of the assay was confirmed using samples from 106 wild mammals that were previously subject to Leishmania sp. kDNA analysis through singleplex reactions. PRINCIPAL FINDINGS: The following optimal conditions were established: 95°C for 1 min followed by 30 cycles of 95°C for 30 s, 61°C for 30 s, and 72°C for 30 s and a final extension at 72°C for 1 min. The multiplex PCR system was capable of detecting 0.1 ng of L. tropica diluted in 100 ng of mammalian DNA. Of 51 kDNA samples that were previously found to be positive, 45 (88.2%) were positive for both targets, two were positive only for kDNA and four were negative for both. Of 55 kDNA samples that were previously identified as negative, 38 (69.1%) were positive for gapdh whereas the other 17 were negative for both targets. CONCLUSIONS/SIGNIFICANCE: The proposed multiplex PCR system allows the simultaneous detection of the gapdh gene and Leishmania sp. kDNA in tissue samples derived from distinct wild mammal species. The amplification of the gapdh mammalian gene in the same reaction ensures the quality and viability of the DNA in the sample, eliminating the possibility of false-negative results that would impair an accurate description of the infection rates in a given population.

First description of Leishmania (Viannia) infection in Evandromyia saulensis, Pressatia sp. and Trichophoromyia auraensis (Psychodidae: Phlebotominae) in a transmission area of cutaneous leishmaniasis in Acre state, Amazon Basin, Brazil.

Studies on the sandfly fauna to evaluate natural infection indexes are still limited in the Brazilian Amazon, a region with an increasing incidence of cutaneous leishmaniasis. Here, by using a multiplex polymerase chain reaction directed to Leishmania kDNA and hybridisation, we were able to identify L. (Viannia) subgenus in 12 out of 173 sandflies captured in the municipality of Rio Branco, Acre state, revealing a positivity of 6.94%. By sequencing the Leishmania 234 bp-hsp70 amplified products from positive samples, infection by L. (V.) braziliensis was confirmed in five sandflies: one Evandromyia saulensis, three Trichophoromyia auraensis and one Pressatia sp. The finding of L. (Viannia) DNA in two Ev. saulensis corresponds to the first record of possible infection associated with this sandfly. Moreover, our study reveals for the first time in Brazil, Th. auraensis and Pressatia sp. infected by L. (Viannia) parasites.

First description of Leishmania (Viannia) infection in Evandromyia saulensis, Pressatia sp. and Trichophoromyia auraensis (Psychodidae: Phlebotominae) in a transmission area of cutaneous leishmaniasis in Acre state, Amazon Basin, Brazil

Studies on the sandfly fauna to evaluate natural infection indexes are still limited in the Brazilian Amazon, a region with an increasing incidence of cutaneous leishmaniasis. Here, by using a multiplex polymerase chain reaction directed to Leishmania kDNA and hybridisation, we were able to identify L. (Viannia) subgenus in 12 out of 173 sandflies captured in the municipality of Rio Branco, Acre state, revealing a positivity of 6.94%. By sequencing the Leishmania 234 bp-hsp70 amplified products from positive samples, infection by L. (V.) braziliensis was confirmed in five sandflies: one Evandromyia saulensis, three Trichophoromyia auraensis and one Pressatia sp. The finding of L. (Viannia) DNA in two Ev. saulensis corresponds to the first record of possible infection associated with this sandfly. Moreover, our study reveals for the first time in Brazil, Th. auraensis and Pressatia sp. infected by L. (Viannia) parasites.

Usefulness of real time PCR to quantify parasite load in serum samples from chronic Chagas disease patients.

BACKGROUND: Inconclusive results of serological diagnosis in Chagas disease have an important impact on blood banks worldwide, reflecting in the high number of discarded bags or in an increased transmission by blood transfusion. Molecular techniques such as qPCR have been used for diagnosis and to monitor Trypanosoma cruzi load in peripheral blood samples. A promising perspective refers to the possibility of parasite DNA detection in serum, taking advantage in using the same samples collected for serological screening. METHODS: In order to evaluate the effectiveness of a qPCR strategy for detecting and quantifying T. cruzi DNA in serum, we selected 40 chronic Chagas disease patients presenting different clinical manifestations: Cardiac (23), Digestive (4), Mixed form [cardiodigestive] (7), and asymptomatic (6). Twenty seronegative individuals from non-endemic areas were included as controls. Samples were extracted using QIAamp DNA mini kit (QIAGEN) and qPCR was performed in a multiplex format with TaqMan probes for the nuclear satellite DNA of T. cruzi and for the human RNase P gene. In addition, DNA migration to serum during blood coagulation was assessed using a commercial exogenous control (Exo IPC, Applied Biosystems) in a separate qPCR reaction. RESULTS: The comparative duplex qPCR analysis revealed that, even with an increase in Ct values, it was possible to detect all DNA targets in serum. In addition, the same linearity range for T. cruzi quantification (from 10(5) to 0.5 par. eq./mL) between serum, blood or culture samples (T. cruzi epimastigotes - Cl Brener strain) was found. When patient samples were evaluated, no significant differences in parasite load between the distinct clinical manifestations were found for both blood and serum samples. Moreover, median values of parasite burden were 1.125 and 1.230 par. eq./mL for serum and blood, respectively. Using serology as gold standard, we found 95% sensitivity for T. cruzi detection in serum and 97.5% for blood, and 100% specificity for both samples. CONCLUSIONS: Taken together, our data indicate the potential of using serum samples for molecular diagnosis and parasite load quantification by qPCR, suggesting its use in reference laboratories for the diagnosis of Chagas disease patients.

Quantificação de DNA de Trypanosoma cruzi em soro e potencial uso das 5 UTRs da família gênica de trans-sialidases para a genotipagem do parasito como complemento ao diagnóstico molecular da infecção / Trypanosoma cruzi DNA quantitation in serum and potential use of 5 RTUs gene family of trans-sialidase for genotyping parasite in addition to molecular diagnosis of infection

Ensaios de PCR para o diagnóstico e monitoramento da carga parasitária em pacientes acometidos pela doença de Chagas são realizados a partir de DNA recuperado de sangue periférico. Uma perspectiva seria a possibilidade do uso de soro para a detecção e quantificação de DNA de T. cruzi, aproveitando amostras coletadas para triagem sorológica em laboratórios de referência e bancos de sangue. Outra limitação é que o diagnóstico molecular com os alvos mais comumente utilizados (kDNA, DNA Satélite) não é capaz de identificar as linhagens do parasito. Tem sido relatada uma baixa sensibilidade para alguns marcadores de genotipagem de T. cruzi quando aplicados diretamente em sangue de pacientes crônicos, uma vez que são cópias únicas ou distribuídos embaixa frequência no genoma do parasito. Estudos baseados em famílias multigênicas, como as proteínas de superfície (trans-sialidases), sugerem que estas sejam capazes de reunir cepas/isoladosde T. cruzi em grupos biologicamente distintos, visando sua caracterização. A presença de sítiospolimórficos nas regiões não traduzidas (UTRs) dos genes de trans-sialidases (TS), além de representar uma estratégia de sobrevivência do parasito ao estresse ambiental, propicia o uso destas regiões como potenciais marcadores moleculares para tipagem de T. cruzi. Este trabalho avaliou o potencial uso do soro e do segmento 5’ UTR de TS de T. cruzi, em ensaios de quantificação de DNAe genotipagem do parasito, respectivamente. Neste sentido, empregamos a PCR em Tempo Real(qPCR) multiplex para a detecção/quantificação de T. cruzi em amostras pareadas de soro e sangue de 40 pacientes com a doença crônica. Para avaliar o uso da 5’ UTR de TS como marcador molecular de genotipagem, utilizamos um painel de cepas/clones de T. cruzi, representantes das seis DTUs (Unidades Discretas de Tipagem)...

PCR-based assays for molecular diagnosis and to evaluate Trypanosoma cruzi load inpatients with Chagas disease are usually performed with DNA recovered from peripheralblood samples. One promising approach would be the use of patients’ serum for thedetection and quantification of T. cruzi DNA, as it would make possible to take advantage ofthe same samples collected for serological screening in blood banks and referencelaboratories. Another limitation is that molecular diagnosis with the most common moleculartargets (kDNA and Satellite DNA) is not suitable for identifying the infecting parasite strain. Alow sensitivity of some T. cruzi genotyping markers has been reported, when those areapplied directly to blood of chronic patients, since they are represented as a single copy orpresent low frequencies in the parasite genome. Studies based on multigene families, suchas the surface proteins (trans-sialidase), suggested the ability of those genes to clusterstrains/isolates of T. cruzi into biologically distinct groups. The presence of polymorphic sitesin the untranslated regions (UTRs) of trans-sialidase genes (TS), as well as represents aparasite’s survival strategy to environmental stress, it promotes the use of these regions aspotential molecular markers for T. cruzi typing. This study evaluated the potential use ofserum and the segment 5' UTR of TS, as new approaches for DNA quantification and T.cruzi genotyping, respectively. We used multiplex Real-Time PCR (qPCR) fordetecting/quantifying parasites in serum and blood paired samples from 40 chronic Chagasdisease patients. In order to investigate the potential use of 5' UTR of TS sequences asmolecular markers for genotyping, we used a panel of T. cruzi strains/clones, representativeof the six DTUs (Discrete Typing Units)...

Preceptoria de enfermagem na residência multiprofissional em oncologia: um estudo descritivo / Nursing preceptorship in multi-professional residency in oncology: a descriptive study / Preceptoria de enfermería en residencia multiprofesional en oncología: un estudio descriptivo

OBJETIVO: Analisar a percepção de residentes sobre fatores potencializadores e limitantes da preceptoria em um programa de residência multiprofissional em oncologia. MÉTODO: estudo descritivo de abordagem qualitativa, com aplicação da metodologia da problematização e análise de conteúdo. RESULTADOS: a participação foi intensa e reafirmou a preceptoria como questão prioritária para o grupo, discutindo-a sob múltiplas dimensões.DISCUSSÃO: A preceptoria exerce papel fundamental no processo de construção de conhecimento significativo na formação humana e profissional dos residentes, requerendo de seus atores um olhar diferenciado para perceber a dinamicidade deste processo. CONCLUSÃO: A percepção dos residentes sobre preceptoria revela a necessidade da qualificação pedagógica para profissionais que realizam esta atividade. Apesar das críticas, facilitadas pela aplicação da problematização, na visão dos participantes, o saldo de aprendizagem no programa de residência é positivo.

AIM: To analyze the perception of resident nurses in terms of the potentiating and limiting factors related to preceptorship in a multidisciplinary residency program in oncology. METHOD: This is a descriptive study that uses a qualitative approach. The methodology of problematization and content analysis was applied in this study. RESULTS: Participation was intense, and preceptorship was reaffirmed as a priority for the group, by means of a multi-dimensional discussion. DISCUSSION: Preceptorship plays a fundamental role in the meaningful knowledge building process in terms of the human and professional training of resident nurses, requiring of its actors a different perspective to perceive the dynamics of the process. CONCLUSION: The perception of residents in terms of preceptorship reveals the need for teaching qualifications for the professionals involved in this activity. Despite the criticism related to the application of the problematization methodology, in the view of the participants, the learning resulting from the residency program is positive.

OBJETIVO: Analizar la percepción de residentes sobre factores potencializadores y limitantes de la preceptoria en un programa de residencia multiprofesional en oncología. MÉTODO: estudio descriptivo de enfoque cualitativo, con aplicación de la metodología de la problematización y análisis de contenido. RESULTADOS: la participación fue intensa y reafirmó la preceptoria como una cuestión prioritaria para el grupo, discutiéndola sobre múltiplas dimensiones. DISCUSIÓN: La preceptoria ejerce un papel fundamental en el proceso de construcción de conocimiento significativo en la formación humana y profesional de los residentes, requeriendo de sus actores una forma de ver diferente para entender la dinámica de este proceso. CONCLUSIÓN: La percepción de los residentes sobre preceptoria revela la necesidad de la calificación pedagógica para profesionales que realizan esta actividad. A pesar de las críticas, facilitadas por la aplicación de la problematización, en la visión de los participantes, el saldo de aprendizaje en el programa de residencia y positivo.

[Problematizing the multidisciplinary residency in oncology: a practical teaching protocol from the perspective of nurse residents]. / Problematizando a residência multiprofissional em oncologia: protocol de ensino prático na perspective de residents de enfermagem.

OBJECTIVE: To investigate practical teaching of nurse residents in a multidisciplinary residency in oncology. METHOD: A qualitative descriptive study grounded in the problematization methodology and its steps, represented by the Maguerez Arch. Data were analyzed using content analysis. RESULTS: Potentiating and limiting elements of the residency guided the design of a practical teaching protocol from the perspective of residents, structured in three stages: Welcoming and ambience; Nursing care for problem situations; and, Evaluation process. CONCLUSION: Systematization of practical teaching promoted the autonomy of individuals and the approximation of teaching to reality, making residency less strenuous, stressful and distressing.

Problematizing the multidisciplinary residency in oncology: a practical teaching protocol from the perspective of nurse residents / Problematizando la residencia multiprofesional en oncología: protocolo de enseñanza práctica desde la perspectiva de residentes de enfermería / Problematizando a residência multiprofissional em oncologia: protocolo de ensino prático na perspectiva de residentes de enfermagem

Objective: To investigate practical teaching of nurse residents in a multidisciplinary residency in oncology. Method: A qualitative descriptive study grounded in the problematization methodology and its steps, represented by the Maguerez Arch. Data were analyzed using content analysis. Results: Potentiating and limiting elements of the residency guided the design of a practical teaching protocol from the perspective of residents, structured in three stages: Welcoming and ambience; Nursing care for problem situations; and, Evaluation process. Conclusion: Systematization of practical teaching promoted the autonomy of individuals and the approximation of teaching to reality, making residency less strenuous, stressful and distressing.

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Objetivo: Investigar la enseñanza práctica de los residentes de enfermería de una residencia oncológica multiprofesional. Método: Investigación descriptiva cualitativa, basada en la metodología de problematización y en sus etapas, representadas por el Arco de Maguerez. Los datos fueron analizados mediante el análisis de contenido. Resultados: Tanto factores potenciadores y limitantes de la residencia guiaron el diseño de un protocolo de enseñanza práctica desde la perspectiva de los residentes, estructurado en tres etapas: acogida y ambiente; cuidados de enfermería a partir de situaciones problema y proceso de evaluación. Conclusión: La sistematización de la enseñanza práctica promueve la autonomía de los sujetos y la aproximación de la enseñanza a la realidad, haciendo la residencia menos agotadora, angustiante y estresante. 
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Objetivo: Investigar o ensino prático de residentes de Enfermagem de uma residência oncológica multiprofissional. Método: Pesquisa descritiva qualitativa, embasada na metodologia da problematização e em suas etapas, representadas pelo Arco de Maguerez. Os dados foram analisados segundo a análise de conteúdo. Resultados: Elementos potencializadores e limitantes da residência nortearam o desenho de um protocolo de ensino prático na perspectiva dos residentes, estruturado em três etapas: acolhimento e ambiência; cuidados de Enfermagem a partir de situações-problema e processo de avaliação. Conclusão: A sistematização do ensino prático promoveu a autonomia dos sujeitos e a aproximação do ensino à realidade, tornando a residência menos extenuante, aflitiva e estressante. 
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Towards the establishment of a consensus real-time qPCR to monitor Trypanosoma cruzi parasitemia in patients with chronic Chagas disease cardiomyopathy: a substudy from the BENEFIT trial.

Quantitative real-time PCR (qPCR) is an accurate method to quantify Trypanosoma cruzi DNA and can be used to follow-up parasitemia in Chagas disease (CD) patients undergoing chemotherapy. The Benznidazole Evaluation for Interrupting Trypanosomiasis (BENEFIT) study is an international, multicenter, randomized, double-blinded and placebo-controlled clinical trial to evaluate the efficacy of benznidazole (BZ) treatment in patients with chronic Chagas cardiomyopathy (CCC). One important question to be addressed concerns the effectiveness of BZ in reducing overall parasite load in CCC patients, even in the absence of parasitological cure. This report describes the evaluation of multiple procedures for DNA extraction and qPCR-based protocols aiming to establish a standardized methodology for the absolute quantification of T. cruzi DNA in Guanidine-EDTA blood (GEB) samples. A panel of five primer sets directed to the T. cruzi nuclear satellite DNA repeats (Sat-DNA) and to the minicircle DNA conserved regions (kDNA) was compared in either SYBR Green or TaqMan systems. Standard curve parameters such as, amplification efficiency, coefficient of determination and intercept were evaluated, as well as different procedures to generate standard samples containing pre-established T. cruzi DNA concentration. Initially, each primer set was assayed in a SYBR Green qPCR to estimate parasite load in GEB samples from chronic Chagas disease patients. The results achieved from Bayesian transmutability analysis elected the primer sets Cruzi1/Cruzi2 (p=0.0031) and Diaz7/Diaz8 (p=0.0023) coupled to the QIAamp DNA Kit extraction protocol (silica gel column), as the most suitable for monitoring parasitemia in these patients. Comparison between the parasite burden of 150 GEB samples of BENEFIT patients from Argentina, Brazil and Colombia, prior to drug/placebo administration, was performed using Cruzi1/Cruzi2 primers in a SYBR Green approach. The median parasitemia found in patients from Argentina and Colombia (1.93 and 2.31 parasite equivalents/mL, respectively) was around 20 times higher than the one estimated for the Brazilian patients (0.1 parasite equivalents/mL). This difference could be in part due to the complexity of T. cruzi genetic diversity, which is a factor possibly implicated in different clinical presentations of the disease and/or influencing parasitemia levels in infected individuals from different regions of Latin America. The results of SYBR Green qPCR assays herein presented prove this methodology to be more cost efficient than the alternative use of internal fluorogenic probes. In addition, its sensitivity and reproducibility are shown to be adequate to detect low parasitemia burden in patients with chronic Chagas disease.

A residência como cenário educativo para enfermeiros: o uso da metodologia da problematização / The residency as an educational scenario for nurses: the use of the problematization methodology

A aplicação da metodologia da problematização no processo de ensino-aprendizagem e nas relações estabelecidas durante as atividades de uma residência pode contribuir para que o aluno se descubra enquanto ser integral e como parte de uma totalidade, resgatando seus conhecimentos e aplicando-os à realidade, aprendendo assim de modo significativo. Isto possibilita formar profissionais mais preparados para o trabalho em equipe e para a integralidade da atenção à saúde, capazes de continuar a aprender durante a vida profissional (AGUILAR-DA-SILVA; ROCHA JUNIOR, 2010). OBJETIVO: Primário - Analisar a aplicação da metodologia da problematização no ensino de residentes de Enfermagem de um Programa de Residência Multiprofissional em Oncologia. Secundários - Identificar as dimensões de problemas vivenciados pelos residentes de Enfermagem de um Programa de Residência Multiprofissional em Oncologia; Discutir possíveis avanços na construção do conhecimento sobre os problemas vivenciados pelos residentes de Enfermagem de um Programa de Residência Multiprofissional em Oncologia com base na aplicação da metodologia da problematização. METODOLOGIA: Trata-se de uma pesquisa descritiva com abordagem qualitativa, desenvolvida em um hospital ensino da rede pública do Rio de Janeiro, que oferece o Programa de Residência Multiprofissional em Oncologia, onde os participantes da pesquisa foram residentes de Enfermagem deste programa, ingressos no ano de 2012. O projeto foi submetido e aprovado por Comitê de Ética reconhecido no CNS, registro CAAE - 0062.0.153.000-10, processo nº 1236/2010. Desta forma segue todos os quesitos éticos. A coleta de dados foi embasada nas etapas do arco de Maguerez (observação da realidade ­ pontos-chave ­ teorização ­ hipóteses de solução ­ aplicação à realidade), e posteriormente analisados segundo Bardin. RESULTADOS: Os dados foram analisados em duas categorias temáticas: A prática assistencial da Enfermagem como cenário no processo ensinoaprendizagem e Metodologia da Problematização: estratégia de ensino para a autonomia do residente de Enfermagem. Quanto aos participantes percebeu-se um grupo jovem, com pouco tempo de formado e possível inexperiência profissional enquanto enfermeiros, entretanto participativo, interessado e disposto a discutir questões de seu cotidiano, onde trouxeram como base para o debate a situaçãoproblema "A desestrutura da residência com foco na preceptoria". O grupo como um todo levantou questionamentos abrangendo várias dimensões do problema, desde sua concepção conceitual, histórica, perpassando a jurídica, operacional, econômica até a emocional/afetiva, além de construírem soluções ou pelo menos preposições para solução do mesmo. Durante o debate, os participantes identificaram também fatores potencializadores e limitantes da preceptoria no programa em questão. PRODUTO: Tendo como iminente anseio transmitido pelo grupo, seu acolhimento e treinamento no cenário prático por parte dos preceptores e equipe de saúde em geral, este estudo traz como produto e contribuição à construção de um protocolo de ensino prático do novo residente no cenário da residência, partindo desde seu acolhimento e ambiência, perpassando o planejamento e implementação dos cuidados de Enfermagem a partir de situações-problema até a avaliação de todo o processo. CONCLUSÃO: A partir da aplicação da problematização, percebeu-se o acréscimo de conhecimento do grupo, advindo do potencial para o estímulo à autonomia dos sujeitos, onde os mesmos aprendem por meio da investigação e reflexão crítica de seus objetivos de aprendizagem, relacionando-os a realidade e por conseguinte, transformando-a. Assim, a metodologia da problematização emerge como uma estratégia pedagógica, por aproximar o ensino à realidade, onde indivíduos de diferentes contextos sociais interagem intercambiando saberes e experiências, ressignificando conceitos e valores, refletindo sobre a realidade, repensando pontos problemáticos, pesquisando, questionando, teorizando e buscando soluções para alteração, transformação da realidade vivida, através da reflexão, mas principalmente da ação

The application of the methodology of problematization in the teaching - learning and the relationships established during the activities of a residence can contribute to the student to discover while being full and as part of a whole, rescuing their knowledge and applying them to reality, learning so significantly. This enables training professionals better prepared for teamwork and for comprehensive health care, able to continue to learn throughout their professional life (AGUILAR -DA - SILVA; ROCK JUNIOR, 2010). OBJECTIVE: Primary - To evaluate the application of the methodology of problematization in teaching nursing residents from Multidisciplinary Residency Program in Oncology. Secondaries - Identify the dimensions of the problems experienced by nursing residents from Multidisciplinary Residency Program in Oncology; Discuss possible advances in the construction of knowledge about the problems experienced by nursing residents from Multidisciplinary Residency Program in Oncology, based on the application of the methodology of problematization. METHODOLOGY: This was a descriptive with approach qualitative, developed in a public teaching hospital in Rio de Janeiro, offering the Multidisciplinary Residency Program in Oncology, where the participants were nursing residents of this program, has entered in year 2012. The project was approved by the Ethics Committee acknowledged the CNS, record CAAE - 0062.0.153.000-10, process nº 1236/2010. The legal ethical aspects were attended. Collecting data was based on the steps of arc Maguerez (observation of reality - key points - theory - possible solutions - application to reality), and subsequently analyzed according to Bardin, the elaboration of thematic categories. RESULTS: The data were analyzed in two themes: The practice of nursing care as a setting in the teaching- learning and methodology of problematization: teaching strategy for the autonomy of the resident of Nursing. As for the participants noticed a young group, with small training time and possible professional inexperience as nurses, participatory however, interested and willing to discuss issues of their daily lives, which brought as a basis for discussion the problem situation " The destructures the residence with focus in the preceptorship". The group as a whole raised questions covering various dimensions of the problem, since its dimension conceptual, historical, going through the legal, operational, economic to the emotional/affective, and build solutions or at least prepositions for the same solution. During the discussion, participants also identified potential and limiting factors the preceptorship of this program in question. PRODUCT: Having as imminent wish transmitted by the group, their reception and training in the practical setting by the preceptors and health professionals in general, this study brings the product and contribution to the construction of a practical teaching protocol new resident in the scenario of residence, starting from his reception and ambience, passing the planning and implementation of nursing care from problem situations to the evaluation of the whole process. CONCLUSION: From the application of questioning, it was noticed the increase of knowledge of the group, coming from the potential for stimulating the autonomy of individuals, where they learn through research and critical reflection of their learning objectives, linking them reality and thus transforming it. Thus, the problem methodology emerges as a pedagogical strategy for teaching approach to reality, where individuals from different social contexts interact by exchanging knowledge and experiences, redefines concepts and values, reflecting on reality, rethinking trouble spots, researching, questioning, theorizing and seeking solutions for change, transformation of the reality through reflection, but mostly the action

Problematization methodology in nursing of oncologic education: a descriptive study

This is a dissertation for the Professional Masters in Health Education from Federal Fluminense University. Aims: To propose and implement a theoretical and practical education tool based on the problematization methodology in nursing in a Multidisciplinary Residency Program. It is an evaluation of the possible learning strategies adopted by nursing students in the of hospital care practice using the methodology of problematization. In it we discuss the methodology of the problematization and its implications for the teaching and practice of nursing. Methods: A descriptive study with a qualitative approach, undertaken in an oncology hospital in Rio de Janeiro. The subjects are students of the Nursing Residency Program in Oncology of this hospital, and ethical aspects will be in accordance with Resolution 196/96.

Chagas disease in the State of Pernambuco, Brazil: analysis of admissions and mortality time series.

INTRODUCTION: A time series study of admissions, deaths and acute cases was conducted in order to evaluate the context of Chagas disease in Pernambuco. METHODS: Data reported to the Information Technology Department of the Brazilian National Health Service between 1980 and 2008 was collected for regions and Federal Units of Brazil; and microregions and municipalities of Pernambuco. Rates (per 100,000 inhabitants) of hospitalization, mortality and acute cases were calculated using a national hospital database (SIH), a national mortality database (SIM) and the national Information System for Notifiable Diseases (SINAN), respectively. RESULTS: The national average for Chagas disease admissions was 0.99 from 1995 to 2008. Pernambuco obtained a mean of 0.39 in the same period, with the highest rates being concentrated in the interior of the state. The state obtained a mean mortality rate of 1.56 between 1980 and 2007, which was lower than the national average (3.66). The mortality rate has tended to decline nationally, while it has remained relatively unchanged in Pernambuco. Interpolating national rates of admissions and deaths, mortality rates were higher than hospitalization rates between 1995 and 2007. The same occurred in Pernambuco, except for 2003. Between 2001 and 2006, rates for acute cases were 0.56 and 0.21 for Brazil and Pernambuco, respectively. CONCLUSIONS: Although a decrease in Chagas mortality has occurred in Brazil, the disease remains a serious public health problem, especially in the Northeast region. It is thus essential that medical care, prevention and control regarding Chagas disease be maintained and improved.

Chagas disease in the State of Pernambuco, Brazil: analysis of admissions and mortality time series / Doença de Chagas no Estado de Pernambuco, Brasil: análise de séries históricas das internações e da mortalidade

INTRODUCTION: A time series study of admissions, deaths and acute cases was conducted in order to evaluate the context of Chagas disease in Pernambuco. METHODS: Data reported to the Information Technology Department of the Brazilian National Health Service between 1980 and 2008 was collected for regions and Federal Units of Brazil; and microregions and municipalities of Pernambuco. Rates (per 100,000 inhabitants) of hospitalization, mortality and acute cases were calculated using a national hospital database (SIH), a national mortality database (SIM) and the national Information System for Notifiable Diseases (SINAN), respectively. RESULTS: The national average for Chagas disease admissions was 0.99 from 1995 to 2008. Pernambuco obtained a mean of 0.39 in the same period, with the highest rates being concentrated in the interior of the state. The state obtained a mean mortality rate of 1.56 between 1980 and 2007, which was lower than the national average (3.66). The mortality rate has tended to decline nationally, while it has remained relatively unchanged in Pernambuco. Interpolating national rates of admissions and deaths, mortality rates were higher than hospitalization rates between 1995 and 2007. The same occurred in Pernambuco, except for 2003. Between 2001 and 2006, rates for acute cases were 0.56 and 0.21 for Brazil and Pernambuco, respectively. CONCLUSIONS: Although a decrease in Chagas mortality has occurred in Brazil, the disease remains a serious public health problem, especially in the Northeast region. It is thus essential that medical care, prevention and control regarding Chagas disease be maintained and improved.

INTRODUÇÃO: Foi realizado estudo de séries históricas de internações, óbitos e casos agudos por doenças de Chagas objetivando avaliar o contexto desta enfermidade em Pernambuco. MÉTODOS: Foram coletados dados notificados de 1980 a 2008 ao Departamento de Informática do Sistema Único de Saúde (DATASUS/MS) para regiões e unidades federativas do Brasil, microrregiões e municípios pernambucanos. As taxas (por 100.000 habitantes) de internações, mortalidade e casos agudos foram obtidas por consulta ao Sistema de Informações Hospitalares (SIH), Sistema de Informação sobre Mortalidade (SIM) e Sistema de Informações de Agravos de Notificação (SINAN), respectivamente. RESULTADOS: A média de internações nacional por doença de Chagas ficou em 0,99 no período de 1995 a 2008. Pernambuco, neste intervalo, apresentou média de 0,39, com as maiores taxas concentradas no interior do estado. Este estado obteve média de óbitos 1,56 entre 1980 e 2007, valor inferior a brasileira (3,66). O país demonstrou declínio de óbitos na análise de tendência, com Pernambuco encontrando-se em estado estacionário para esta taxa. Interpolando os dados referentes a internações e óbitos, evidenciou-se mortalidade em valores superiores as taxas de internações nacionais, entre 1995 e 2007. O mesmo fato ocorreu em Pernambuco, exceto em 2003. Entre 2001 e 2006, a taxa de casos agudos foi de 0,56 e 0,21 respectivamente para Brasil e para Pernambuco. CONCLUSÕES: Mesmo o Brasil demonstrando redução na mortalidade, a doença permanece como grave problema de saúde pública, principalmente no nordeste. Desta forma, é fundamental a manutenção e melhoria das ações de atenção médica, controle e prevenção já existentes.

Avaliação da reação em cadeia da polimerase na detecção de trypanosoma cruzi em soro e em sangue de pacientes com doença de chagas crônica / Evaluation of polymerase chain reaction detection of trypanosoma cruzi in serum and blood of patients with chronic chagas disease

O diagnóstico sorológico da doença de Chagas apresenta algumas desvantagens, tanto na fase aguda devido à falta de kits comerciais aprovados pela ANVISA, quanto na fase crônica, nos casos de diagnósticos inconclusivos ou discordantes, além de não ser o método ideal para diagnosticar a infecção por transmissão congênita. Uma alternativa para a confirmação é o uso da técnica da PCR que apresenta alta sensibilidade e especificidade. No presente trabalho, objetivou-se avaliar a eficácia da ferramenta da biologia molecular, indicada pelo Ministério da Saúde como diagnóstico confirmatório da doença de Chagas, a PCR, utilizando amostras de soro como um potencial meio de extração de DNA do parasito. Para isso, foram coletadas amostras de sangue e soro de 65 pacientes chagásicos (FC=39, FM=10, FD=5, FI=9 e DES=2), selecionados no Ambulatório de Doença de Chagas do Hospital Universitário Oswaldo Cruz. Todos os pacientes foram avaliados clinicamente para caracterização das formas clínicas e realizaram exames confirmatórios para infecção pelo T. cruzi. Além disso, também foram coletadas amostras de 35 indivíduos não chagásicos (área endêmica=9 e área não endêmica=26) que apresentaram sorologia negativa para infecção. Inicialmente, foram analisadas três formas de extração do material de sangue e soro. Após a escolha da melhor forma de extração, as amostras de sangue e soro dos pacientes chagásicos crônicos e não chagásicos foram aplicados na técnica de PCR com a finalidade de avaliar a sensibilidade e especificidade, como também a concordância dos resultados. Ocorreu predominância da forma cardíaca (60 por cento) entre os portadores da infecção pelo T. cruzi. Os resultados da técnica de PCR, utilizando amostras de soro, apresentaram uma sensibilidade estatisticamente superior (61,3 por cento) à amostra de sangue (41,5 por cento). Quando analisado o resultado utilizando as amostras simultaneamente (sangue e soro), obteve-se um valor da sensibilidade aumentado...

Higienização das mãos: o ensino e a prática entre graduandos na área da saúde / Hand washing: teaching and practice among undergraduate health sector students

Estudo realizado com graduandos do último ano/semestre da área da saúde de Instituições de Ensino Superior (IES) em Goiás/2005, com objetivo de verificar a compreensão destes acerca da higienização das mãos (HM), identificar a contribuição das IES na formação dos alunos sobre esta temática, identificar fatores que favorecem a adesão à HM e verificar a disponibilidade de recursos materiais para HM nos Estabelecimentos de Assistência à Saúde (EAS) utilizados como cenários de prática. Observados os aspectos éticos, os dados foram obtidos pela aplicação de questionário, previamente avaliado, e processados no programa Epiinfo (CDC, 2004), quando indicado, aplicado o teste qui-quadrado. Participaram do estudo 777 graduandos (68,0%); dos cursos de Biomedicina, Enfermagem, Farmácia, Fisioterapia, Fonoaudiologia, Medicina, Odontologia e Terapia Ocupacional. Houve discordância entre o entendimento sobre HM e a adesão. Embora tenhamos evidenciado que as IES tenham abordado este tema durante a formação dos profissionais, isto não foi suficiente para alterar a práxis

This study was undertaken with students during their last year/semester of health sector studies at Higher Education Institutions (HEI), in Goiás State/2005. It aimed to verify the understanding of these students about hands washing (HW), identify the contribution of the HEI to the students' education on this theme, identify factors that favor the practice of HW, and verify the availability of material resources for HW at Health Assistance Establishments (HAE) used as practice scenarios. After observing ethical aspects, the data was obtained through a previously-evaluated questionnaire, and processed using Epi Info software (CDC, 2004), when the chi-square test was applied. 777 students took part (68.0%) in the study, belonging to the following courses: Biomedicine, Nursing, Pharmacy, physiotherapy, phonoaudiology, Medicine, Dentistry and Occupational Therapy. There was a noticeable difference between the understanding on HW and actual practice. Although we evidenced that HEIs have approached this subject during the educational process, this was not enough to modify the practice