Development of Multiplex Polymerase Chain Reaction (PCR)-Based MSA Assay for Bladder Cancer Detection.

Several studies have shown that microsatellite changes can be profiled in the urine to detect bladder cancer. Microsatellite analysis (MSA) of bladder cancer detection requires a comprehensive analysis of up to 15-20 markers based on amplifying and interpreting many individual MSA markers, which can be technically challenging. To develop fast, efficient, standardized, and less costly MSA to detect bladder cancer, we developed three multiplex polymerase chain reaction (PCR) based MSA assays, all of which were analyzed by a genetic analyzer. First, we selected 16 MSA markers based on nine publications. We developed MSA assays based on triplet or three-tube-based multiplex PCR (Triplet MSA assay) using samples from Johns Hopkins University (JHU Sample, first set of samples). In the second set of samples (samples from six cancer patients and fourteen healthy individuals), our Triplet Assay with 15 MSA markers correctly predicted all 6/6 cancer samples to be cancerous and 14/14 healthy samples to be healthy. Although we could improve our report with more clinical information from patient samples and an increased number of cancer patients, our overall results suggest that our Triplet MSA Assay combined with a genetic analyzer is a potentially time- and cost-effective genetic assay for bladder cancer detection and has potential use as a dependable assay in patient care.

Use of high flow nasal cannula in Spanish neonatal units.

INTRODUCTION: The use of high-flow cannula therapy (HFNC) in neonatal units has increased in recent years, but there are no consensus guidelines on its indications and application strategies. Our aim was to know the rate of use of HFNC, their indications and the management variability among Spanish neonatal units. MATERIAL AND METHODS: Twenty-five-question survey for medical and nursing staff. Level II and III units were contacted by phone and sent in Google forms between September 2016 and December 2018. RESULTS: Ninety-seven responses (63.9% medical, 36.1% nursing), from 69 neonatal units representing 15 autonomous communities (87% level of care III; 13% level II). All units except one have HFNC with a humidified and heated system. Their most frequent indications are: non-invasive ventilation weaning (79.4%), bronchiolitis (69.1%), respiratory distress of the term newborn (58.8%), after extubation (50.5%). Minimum flow (1-5 L/min) and maximum flow (5-15 L/min) are variable between units. 22.7% have experienced some adverse effect from its use (9 air leak, 12 nasal trauma). Less than half have an employment protocol, but all the answers agree on the usefulness of national recommendations. CONCLUSIONS: HFNC therapy is widely used in Spanish units, but there is great variability in its indications and strategies of use. National recommendations would be applicable in most units and would allow unifying its use.

Comparison of SARS-CoV-2 detection in Saliva by real-time RT-PCR and RT-PCR/MALDI-TOF Methods

The coronavirus disease 2019 (COVID-19) pandemic has accelerated the need for rapid implementation of diagnostic assays for detection of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in respiratory specimens. While multiple molecular methods utilize nasopharyngeal specimens, supply chain constraints and need for easier and safer specimen collection warrant alternative specimen types, particularly saliva. Although saliva has been found to be a comparable clinical matrix for detection of SARS-CoV-2, evaluations of diagnostic and analytic performance across platforms for this specimen type are limited. Here, we compared two methods for SARS-CoV-2 detection in saliva: the Roche cobas(R) 6800/8800 SARS-CoV-2 real-time RT-PCR Test and the Agena Biosciences MassARRAY(R) SARS-CoV-2 Panel/MassARRAY(R) System. Overall, both systems had high agreement with one another, and both demonstrated high diagnostic sensitivity and specificity when compared to matched patient upper respiratory specimens. We also evaluated the analytical sensitivity of each platform and determined the limit of detection of the Roche assay was four times lower than that of Agena for saliva specimens (390.6 v. 1,562.5 copies/mL). Furthermore, across individual target components of each assay, T2 and N2 targets had the lowest limits of detection for each platform, respectively. Together, we demonstrate that saliva represents an appropriate specimen for SARS-CoV-2 detection in two technologies that have high agreement and differ in analytical sensitivities overall and across individual component targets. The addition of saliva as an acceptable specimen and understanding the sensitivity for testing on these platforms can further inform public health measures for screening and detection to combat the COVID-19 pandemic.

Micropropagação de framboeseira em diferentes concentrações de ferro / Adjustment of the iron concentration in the culture medium to raspberry micropropagation

Objetivou-se otimizar a micropropagação das cultivares 'Heritage' e 'Batum' de framboeseira, ajustando-se a concentração de ferro do meio de cultura. Os tratamentos, avaliados durante a fase de multiplicação dos explantes, consistiram em modificações do meio MS quanto à concentração de ferro (0, 25, 50 e 75mg L-1 de FeSO4.7H2O). Foram realizados três subcultivos de 45 dias, sob condições ambientais de 25±4°C, 20µE m-2 s-1 e fotoperíodo de 16 horas. Utilizou-se delineamento experimental inteiramente casualizado, em um bifatorial (duas cultivares x quatro doses de FeSO4.7H2O), com quatro repetições, sendo as unidades experimentais constituídas por um frasco contendo cinco explantes de 8-10mm com três a quatro gemas. Independentemente da concentração de ferro do meio de cultura, a 'Heritage' apresentou maior número médio de brotos por explante (6,9), proporcionando a produção de 501,1 plantas em três subcultivos, enquanto que a 'Batum' apresentou número de brotos de 6,0 e produção de 306,3 plantas. A concentração ideal de FeSO4.7H2O no meio de cultura para multiplicação in vitro dos explantes foi de 43,4mg L-1, para a 'Heritage', e de 43,1mg L-1, para a 'Batum'.

The aim of this research was to optimize the micropropagation of 'Heritage' and 'Batum' raspberries, adjusting the iron concentration in the culture medium. The treatments were evaluated during the explant multiplication, using MS medium modified in FeSO4.7H2O concentration (0, 25, 50 and 75mg L-1). The experiment was carried out during three subcultures of 45 days under 25±4°C, 20µE m-2 s-1 and photoperiod of 16 hours. The experimental design was a factorial entirely randomized (two cultivars x four FeSO4.7H2O concentrations) with four replications. The experimental unit consisted of a flask with five explants (8-10mm diameter with three to four shoots). Independently of the iron concentration in the culture medium, 'Heritage' showed the highest number of shoots per explant (6.9) providing the production of 501.1 plants in three subcultures, while 'Batum' showed 6.0 shoots per explant and production of 306.3 plants. The best FeSO4.7H2O concentration in culture medium for explant multiplication was 43.4mg L-1 for 'Heritage' and 43.1mg L-1 for 'Batum'.

A inclusão de crianças com necessidades especiais nas aulas de Educação Física na educação infantil / The inclusion of children with special needs on physical education classes at elementary school / La inclusión de niños con necesidades especiales en clases de educación física en la educación infantil

O presente estudo investiga a temática da inclusão de crianças com necessidades especiais na prática pedagógica da Educação Física na Educação Infantil. A metodologia é de corte qualitativo, os participantes do estudo são professores de educação física da rede regular de ensino que possuem crianças com necessidades especiais nas suas aulas da educação infantil. A coleta de informações utilizou instrumentos como entrevistas e observações. Os resultados do estudo são apresentados em categorias: compreensão da inclusão na prática da educação física; experiências relacionais do professor com crianças com necessidades especiais; perspectivas de qualificação profissional; suporte pedagógico para o processo de inclusão. O estudo permite evidenciar que os professores de educação física reconhecem a inclusão como uma necessidade pedagógica, porém a formação continuada e a prática pedagógica carecem de aprendizagens e avanços nessa área

The current research has investigated the inclusion of children with special needs in the pedagogical practice of Physical Education in elementary school. The methodology is qualitative, the subjects involved in the analysis work as Physical Education teachers in the regular school system and they deal with children with special needs in their classes. The data are composed by interviews and observations. The results are divided into four categories: comprehension of inclusion in the practice of Physical Education; the relation between teachers and children with special needs; prospects in the professional development; pedagogical support to the process of inclusion. This study has showed that Physical Education teachers recognize that inclusion is an important issue in the pedagogical practice. It also has showed that teachers' continuous formation and their pedagogical practice must be developed

El presente estudio investiga la temática de la inclusión social de niños con necesidades en la práctica pedagógica de la Educación Física en la educación infantil. La metodología utilizada en este trabajo es cualitativa, y los participantes son profesores de la red regular de la enseñanza que abarca niños con necesidades especiales. La búsqueda por informaciones ocurrió a través de los siguientes instrumentos: entrevistas y observaciones. Los resultados del estudio son presentados en categorías: comprensión de la inclusión en la práctica de educación física; experiencias relacionales del profesor con niños con necesidades especiales; perspectivas de calificación profesional; soporte pedagógico para el desarrollo de la inclusión. El estudio muestra que los profesores de educación física reconocen la inclusión como una necesidad pedagógica, sin embargo hace falta una formación progresiva, y la práctica de enseñanza con más aprendizaje y avanzos

Neurobiologia do transtorno de humor bipolar e tomada de decisão na abordagem psicofarmacológica / Bipolar disorder: neurobiology and therapeutic decision of psychopharmacological treatments

O Transtorno do Humor Bipolar (THB) caracteriza-se por oscilações do humor que causam prejuízos significativos no âmbito biopsicossocial. O interesse da comunidade científica por este transtorno vem aumentando nos últimos cinco anos em função de sua crescente prevalência associada ao refinamento diagnóstico, à ampliação do arsenal terapêutico e ao conhecimento dos avanços nas pesquisas da neurobiologia do transtorno. A presente revisão aborda questões diagnosticas e terapêuticas aplicadas à neurobiologia dos THB, relacionando-as diretamente à terapêutica dos quadros de mania, hipomania, estados mistos, depressão bipolar e ciclagem rápida, da infância à idade adulta. São revisados criticamente importantes estudos realizados com diferentes fármacos potencialmente eficazes como estabilizadores do humor, nos diversos subdiagnósticos do THB. São analisados fármacos, tais como o lítio, anticonvulsivantes, antipsicóticos, benzodiazepínicos, bloqueadores dos canais de cálcio e hormônio tireoideo, bem como as possíveis bases biológicas para seus efeitos terapêuticos. Em síntese, este trabalho aborda os avanços da psicofarmacologia cuja eficácia é comprovada nos subtipos do THB, procurando relacioná-los com a neurobiologia deste transtorno.