Isothermal digital detection of microRNAs using background-free molecular circuit.

MicroRNAs, a class of transcripts involved in the regulation of gene expression, are emerging as promising disease-specific biomarkers accessible from tissues or bodily fluids. However, their accurate quantification from biological samples remains challenging. We report a sensitive and quantitative microRNA detection method using an isothermal amplification chemistry adapted to a droplet digital readout. Building on molecular programming concepts, we design a DNA circuit that converts, thresholds, amplifies, and reports the presence of a specific microRNA, down to the femtomolar concentration. Using a leak absorption mechanism, we were able to suppress nonspecific amplification, classically encountered in other exponential amplification reactions. As a result, we demonstrate that this isothermal amplification scheme is adapted to digital counting of microRNAs: By partitioning the reaction mixture into water-in-oil droplets, resulting in single microRNA encapsulation and amplification, the method provides absolute target quantification. The modularity of our approach enables to repurpose the assay for various microRNA sequences.

Emerging isothermal amplification technologies for microRNA biosensing: Applications to liquid biopsies.

The potential of microRNAs (miRNAs) as biomarker candidates in clinical practice for diagnosis, prognosis and treatment response prediction, especially in liquid biopsies, has led to a tremendous demand for techniques that can detect these molecules rapidly and accurately. Hence, numerous achievements have been reported recently in miRNA research. In this review, we discuss the challenges associated with the emerging field of miRNA detection, which are linked to the intrinsic properties of miRNAs, advantages and drawbacks of the currently available technologies and their potential applications in clinical research. We summarize the most promising nucleic acid amplification techniques applied to the in vitro detection of miRNAs, with a particular emphasis on the state of the art for isothermal alternatives to RT-qPCR. We detail the sensitivity, specificity and quantitativity of these approaches, as well as their potential for multiplexing. We also review the different detection formats to which these chemistries have been adapted, including analog readouts such as real-time monitoring, digital counting based on single-molecule amplification in compartments, and surface-based strategies.

Streamlined digital bioassays with a 3D printed sample changer.

Droplet-based microfluidics has permeated many areas of life sciences including biochemistry, biology and medicine. Water-in-oil droplets act as independent femto- to nano-liter reservoirs, enabling the parallelization of (bio)chemical reactions with a minimum sample input. Among the range of applications spanned by droplet microfluidics, digital detection of biomolecules, using Poissonian isolation of single molecules in compartments, has gained considerable attention due to the high accuracy, sensitivity and robustness of these methods. However, while the droplet throughput can be very high, the sample throughput of these methods is poor in comparison to well plate-based assays. This limitation comes from the necessity to convert independently each sample into a monodisperse emulsion. In this paper, we report a versatile device that performs the quick sequential partitioning of up to 15 samples using a single microfluidic chip. A 3D printed sample rotor is loaded with all samples and connected to a pressure source. Simple magnetic actuation is then used to inject the samples in the microfluidic chip without pressure disruption. This procedure generates monodisperse droplets with high sample-to-sample consistency. We also describe a fluorescent barcoding strategy that allows all samples to be collected, incubated, imaged and analyzed simultaneously, thus decreasing significantly the time of the assay. As an example of application, we perform a droplet digital PCR assay for the quantification of a DNA amplicon from 8 samples in less than 2 hours. We further validate our approach demonstrating the parallel quantification of 11 microRNAs from a human sample using an isothermal nucleic acid amplification chemistry. As an off-chip device, the sample changer can be connected to a variety of microfluidic geometries and therefore, used for a wide range of applications.

Effect of Hospital Accreditation Process in Outcomes of Patients with Acute Coronary Syndrome

Abstract Background: Hospital accreditation has as goal the standardization of patient care, aiming quality improvement. On 2015, a cardiology reference hospital was evaluated and got level 3 from ONA in care given to Acute Coronary Syndrome (ACS) patients. Objetive: To compare length of stay (LOS) at Coronary Care Unit (CCU) and the total LOS at the hospital of ACS patients before and after ONA 3 accreditation. Other clinical outcomes were also analyzed. Methods: Systematic and prospective registry of admitted ACS patients at CCU, whose population was divided into pre-accreditation (period 1) and post-accreditation (period 2). Descriptive analysis was performed. For statistical analysis the Mann-Whitney test, chi-squared, Fisher's exact test and Multiple Linear Regression were performed. P value was considered statistically significant when < 0,05. Results: 372 patients were admitted with ACS, 186 in period 1, of which 47 (25,3%) with ST segment Elevation Myocardial Infarction (STEMI), and 186 in period 2, of which 70 (37,6%) with STEMI. The mean age was 65,9 years (± 12,2). About the CCU LOS, there was a reduction from 3 (IQR: 2-4) to 2,5 days (IQR: 2-4; p value = 0,088). Regarding the hospital LOS, there was also a reduction from 8 (IQR: 5-12,25) to 6 days (IQR:4-11; p value = 0,004). Analyzing the type of ACS, there was a significant reduction only at the hospital LOS in non-STEMI patients: 8 to 6 days (p value = 0,001). Other hospitalization length of stay and clinical outcomes did not present a significant reduction in the comparison. Conclusion: After the ONA 3 accreditation, there was a reduction of hospital LOS. There were no significant differences in the other outcomes analyzed.

Conhecimento, comportamento e práticas em saúde do homem em relação ao câncer de próstata / Knowledge, behaviour and health practices of men concerning the prostate cancer / Conocimiento, comportamiento y práctica en salud del hombre en relación con cáncer de próstata

Objetivo: Descrever o conhecimento, o comportamento e as práticas em relação ao câncer de próstata em adultos. Métodos: Estudo quantitativo, realizado com 130 homens. A coleta de dados ocorreu no primeiro semestre de 2015, por meio do Modelo de Crenças em Saúde. Resultados: Embora relatem conhecimento sobre o câncer, 30% realizam a dosagem do Antígeno Prostático Específico e 17,6% o exame de toque retal anualmente. A maioria percebe a susceptibilidade e a severidade do câncer e acredita que é capaz de fazer algo por si mesmo, beneficiando-se com esses cuidados. Quanto às barreiras, 16,9% apresentaram comportamento não preventivo. Conclusão: O medo da dor, a vergonha e a falta de coragem para realizar o exame constituem barreiras a esses indivíduos. A equipe de saúde deve oportunizar abordagens considerando esses aspectos, utilizando estratégias que ampliem o acesso dos homens ao serviço de saúde e estimulem a prática do autocuidado

Objective: The study's purpose has benn to describe the knowledge, behavior and health practices regarding the prostate cancer in adults. Methods: It is a cross-sectional and descriptive study with a quantitative approach, which had 130 participating men. Data collection took place from March to April 2015, using the Health Belief Model. Results: Although they report having knowledge about cancer, only 30% had done the prostate-specific antigen dosage and 17.6% the rectal examination annually. Most comprehend the susceptibility and severity of this cancer, and believe that they are capable of doing something for themselves and benefiting from such care. Regarding the barriers, 16.9% had non-preventive behavior. Conclusion: Fear of pain, shame, and lack of courage to take the exam are barriers to these individuals. The health team should opportunistically approach these aspects by using strategies that increase men's access to health care and encourage self-care practices

Objetivo: Describir el conocimiento, el comportamiento y las prácticas en relación al cáncer de próstata en adultos. Métodos: Estudio cuantitativo, realizado con 130 hombres. La recolección de datos ocurrió entre marzo a abril de 2015, a través del Modelo de Creencias en Salud. Resultados: Aunque reportan conocimiento sobre el cáncer, el 30% realiza la dosificación del Antígeno Prostático y el 17,6% el examen de tacto anual. La mayoría percibe la susceptibilidad y la severidad del cáncer y cree que es capaz de hacer algo por sí mismo y beneficiarse con esos cuidados. En cuanto a las barreras, el 16,9% presentó comportamiento no preventivo. Conclusión: El miedo al dolor, la vergüenza y la falta de coraje para realizar el examen constituyen barreras a esos individuos. El equipo de salud debe oportunizar enfoques considerando estos aspectos, utilizando estrategias que amplíen el acceso al servicio de salud y estimulen la práctica del autocuidado

Análise da força de preensão manual e risco cardiovascular de adolescentes com Diabetes Melitos tipo 1 / Analysis of handgrip muscle strength and cardiovascular risk in adolescents with diabetes mellitus type 1

Comparar a força de preensão manual de adolescentes com e sem diabetes mellitus tipo 1 (DM1) e correlacionar as variáveis hemodinâmicas com o controle da glicemia pela hemoglobina glicada (HbA1c) nos adolescentes diabéticos. Método: Foram avaliados 49 adolescentes com DM1 (12,73 ± 1,23 anos; índice de massa corporal 19,52 ± 2,62 kg/m2 ) e 75 adolescentes sem DM1 (13,31 ± 1,16 anos; índice de massa corporal 20,79 ± 3,64 kg/m2 ). A HbA1c foi determinada por cromatografia líquida de alta ficiência (HPLC) e a força de preensão foi obtida com o dinamômetro Jamar®. As variáveis hemodinâmicas registradas foram à frequência cardíaca e a pressão arterial (PA) na posição sentada. Resultados: Os adolescentes do gênero masculino com DM1 apresentaram menores valores de força de preensão manual absoluta da mão direita (26,48 ± 6,24 vs 32,59 ± 9,59 kg; p = 0,004), da mão esquerda (25,45 ± 6,52 vs 30,76 ± 8,19 kg; p = 0,006) e maior PA diastólica (66,43 ± 8,62 vs 72,40 ± 10,01 mmHg; p = 0,019) do que os adolescentes sem DM1. As adolescentes do gênero feminino com DM1 apresentaram menores valores de força de preensão manual absoluta da mão direita (26,20 ± 4,09 vs 29,53 ± 5,27 kg; p = 0,017) e da mão esquerda (24,50 ± 4,29 vs 27,79 ± 5,11 kg; p = 0,017). Houve correlação positiva da frequência cardíaca (0,44; p = 0,01) e da PA diastólica (0,37; p = 0,01) com a HbA1c. Conclusão: Adolescentes com DM1 devem receber atenção quanto a menor força muscular e risco cardiovascular com o aumento da HbA1c...(AU)

To compare the handgrip muscle strength of adolescents with and without diabetes mellitus type 1 (DM1) and to correlate the hemodynamic variables with the glycemic control by glycated hemoglobin (A1c) in the diabetic adolescents. Method: 49 adolescents with DM1 (12.73 ± 1.23 years; body mass index 19.52 ± 2.62 kg/m2 ) and 75 adolescents without DM1 (13.31 ± 1.16 years; body mass index 20.79 ± 3.64 kg/m2 ) were evaluated. The A1c was determined by high performance liquide chromatography (HPLC) and handgrip muscle strength was obtained by the Jamar® dynamometer. The hemodynamic variable measured were heart rate and blood pressure (BP) in the seated position. Results: The male adolescents with DM1 presented lower values of handgrip muscle strength in the right hand (26.48 ± 6.24 vs 32.59 ± 9.59 kg; p = 0.004), and left hand (25.45 ± 6.52 vs 30.76 ± 8.19 kg; p = 0.006) and higher diastolic BP (66.43 ± 8.62 vs 72.40 ± 10.01 mmHg; p = 0.019) as compared with the male adolescents without DM1. The female adolescents with DM1 presented lower values of handgrip strength in the right hand (26.20 ± 4.09 vs 29.53 ± 5.27 kg; p = 0.017), and left hand (24.50 ± 4.29 vs 27.79 ± 5.11 kg; p = 0.017). There was a positive correlation of heart rate (0.44; p = 0.01) and diastolic BP (0.37; p = 0.01) with A1c. Conclusion: Adolescents with DM1 should receive attention regarding their reduced muscle strength and cardiovascular risk with the increase in A1c...(AU)

De quem é essa música? Influência social nas atitudes perante avaliações artísticas / Who wrote this song? Social influence on attitudes concerning artistic evaluations

Este estudo analisou a forma como as pessoas percebem e avaliam uma letra de música quando sua composição é atribuída a um cantor e compositor de renome nacional, regional ou desconhecido, assim como quando ela é atribuída a um homem ou uma mulher. Uma amostra de 168 indivíduos com nível superior completo (84 homens, 84 mulheres) relatou suas atitudes perante letras de músicas fictícias atribuídas a diferentes cantores, avaliando-a em escalas relativas ao quanto gostaram da letra, quão bem escrita a consideraram e quão interessados estariam em ouvi-la. Resultados apontam que, quando associada a cantores nacionais, a letra da música é mais bem avaliada que quando associada a cantores regionais e cantores desconhecidos, respectivamente. O sexo do cantor não influenciou nos julgamentos. Esses resultados são discutidos à luz de alguns modelos de atitude e influência social. (AU)

This study examined people's reaction on the evaluation of song lyrics when they are led to believe the lyrics were written either by a national, regional or unknown singer and composer, as well as when the lyrics are associated to a male or female singer. 168 people with college degrees (84 male, 84 female) were asked to describe their attitudes towards fictitious lyrics in terms of how much they liked it, how well-written they judged it, and how interested they would be in listening to it. Results indicate that lyrics associated to national singers were more favorably evaluated than lyrics associated to regional singers and unknown singers, respectively. The singer's gender did not influence the judgments. These results are discussed in terms of some attitude and social influence models.(AU)

De quem é essa música?Influência social e avaliação artística / Who wrote this song? Social influence and artistic evaluation

Essa pesquisa analisou a forma como o público percebe e avalia uma letra de música quando sua composição é atribuída a um cantor e compositor nacional,regional ou desconhecido, assim como quando ela é atribuída a um homem ou uma mulher. 150 estudantes universitários (73 homens, 77 mulheres) avaliaram a letra de uma música fictícia em termos de quanto gostaram dela, quão bem escrita acharam que ela foi, e se estariam interessados em ouvi-la. Os resultados apontam que letras de música associadas a cantores regionais tiverem uma melhor avaliação que aquelas associadas a cantores desconhecidos, enquanto que as associadas a cantores nacionais não se diferenciaram dos demais grupos.O sexo do cantor não influenciou nos julgamentos. Outro achado foi a tendência das mulheres a gostar mais da letra da música que os homens. Esses resultados são discutidos à luz de alguns modelos de atitude e influência social. (AU)

This research examined the public’s reaction on the evaluation of lyrics when they’re led to believe the lyrics were written either by a national, regional or unknown singer and composer, as well as when the lyrics are associated to amale or female singer. 150 college students (73 male, 77 female) were askedto evaluate fictitious lyrics in terms of how much they liked it, how well-written they judge it, and if they were interested in listening to it. Results indicated that lyrics associated to regional singers were more favorably evaluated than lyrics associated to unknown singers, while those associated to national singers did not achieve a significant difference between any of the other groups. The singer’s sex did not influence the judgments. Other finding was the tendency for women to like the lyrics more than men. These results are discussed in termsof some attitude and social influence models.(AU)

Estudo comparativo de informação em saúde bucal entre estudantes de cursos da área da saúde / Comparative study of oral health information between graduation students of health courses

Este trabalho teve com objetivo comparar o grau de informação em saúde bucal entre os estudantes da área de saúde da Universidade Estadual da Paraíba. Utilizou-se uma abordagem indutiva, com procedimento estatístico descritivo e técnica de documentação direta através da pesquisa de campo, utilizando o formulário comoinstrumento. A amostra foi composta por 200 estudantes dos cursos de Enfermagem, Farmácia, Fisioterapia e Odontologia, divididos em 4 grupos. dentre os principais resultados, observou-se que 76,5 porcento pertenciam ao gênero feminino, com média de idade de 21,7 anos. Com relação à frequência da visita ao cirurgião -dentista, 45,5 porcento compareceram anualmente ao consutório. Para 57,5 porcento dos estudantes, a última consulta odontológica ocorreu de 1 a 6 meses. Quanto à frequência da escovação, 92,5 porcento a realizavam três vezes ou mais ao dia e 65 porcento dos estudantes consideraram sua saúde bucal satisfatória

Atresia do esôfago no recém-nascido: utilizaçäo de Sistema de Aspiraçäo das Vias Aéreas com Baixa Pressäo (Sistema Venturi): estudo de caso / Esophageal atresia in a newborn: the use of a upper airway low pressure aspiration system (Ventury System): case study

O presente estudo de caso relata a utilizaçäo do sistema de aspiraçäo contínua de vias aéreas superiores com baixa pressäo (Sistema Venturi) em um recém-nascido com atresia do esôfago, internado na unidade de cirurgia pediátrica de um hospital na cidade de Säo Paulo...