Destros e canhotos diferem quando comparados em algumas tarefas motoras, parecendo os canhotos usufruir de alguma vantagem em tarefas visuo-motoras. Neste estudo foi analisado, em cada grupo de preferência manual, o efeito da velocidade do estímulo, do sexo e da mão de execução no desempenho de uma tarefa simples de antecipação-coincidência. Participaram 12 destros e 12 canhotos de ambos os sexos, estudantes universitários de Desporto. Empregou-se o "Bassin Anticipation Timer" para avaliar a capacidade de antecipação-coincidência em três velocidades: 268 cm/s, 402,3 cm/s e 536,4 cm/s (6, 9 e 12 mph, respectivamente). Os sujeitos executaram a tarefa tanto com a mão preferida como com a mão não preferida. Principais resultados: 1) apenas os destros foram afetados pela variável velocidade do estímulo, apresentando antecipação das respostas e maior variabilidade na velocidade 268 cm/s, enquanto nas velocidades 402,3 cm/s e 536,4 cm/s as respostas foram enviesadas no sentido do atraso da resposta e com variabilidade menos acentuada na velocidade mais alta; 2) o sexo teve um efeito significativo apenas nos canhotos, sendo o sexo masculino mais preciso e menos enviesado nas suas respostas do que o sexo feminino; 3) a assimetria manual manifestou-se apenas nos canhotos na velocidade de 268 cm/s e no Erro Variável. Concluímos que cada grupo de preferência manual parece comportar-se de forma diferenciada em tarefas perceptivas de Antecipação-Coincidência onde a velocidade do estímulo é manipulada.
The serological monitoring of SARS-CoV-2 infection using the measurement of IgG is an important tool to determine the number of infected people in a population, and the disease spreading. One of the most important proteins from SARS-CoV-2 applied to immune-based assays is the nucleocapsid protein, also known as N protein. This is an abundant protein in the pathogen and exerts important functions in viral viability and replication. Moreover, protein N is highly immunogenic. The use of N protein as a recombinant protein is difficult since this biomolecule is mainly produced inside inclusion bodies in Escherichia coli. In this work, we present a simple and easy method for the expression and purification of soluble his-tagged N protein, suitable for serological assays, such as ELISA. ELISA assays using this recombinant protein presented 100% of specificity and 86% of sensitivity. The protocol for expression and purification of this recombinant protein can be applied in low infrastructure laboratories, without equipment dedicated to protein expression. This work may help other research groups to develop serological assays to monitor antibody production against SARS-CoV-2.
Introduction:The pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has presented an urgency for neutralizing assays. The reference assay, plaque reduction neutralization test (PRNT), is not safe and requires a biosafety level-3 laboratory, since it demand for live viruses. Moreover, PRNT is laborious, time consuming and is not adapted to high throughput, a feature necessary to screen a high number of plasms, small molecules and antibodies.Objectives: Our aim was to develop a safe, easy and faster neutralizing antibodies (nAbs) assay.Methodology: Lentiviral and non-replicative particles (pseudovirus) were developed in HEK293T cells and used to transduce HEK293T-ACE2 overexpressing cells. The automated image analysis was standardized in a high content system, in which the average ZsGreen fluorescence intensity was used to select transduced and non-transduced cells, while the percentage of ZsGreen cells was the readout to calculate the potency of human plasmas (pNT50). Plasma samples from 29 healthy individuals were collected before and after vaccination. Results:The pseudovirus produced was specific for ACE2 overexpressing cells. It was observed a linear correlation between the percentage of ZsGreen + cells and ZsGreen average fluorescence intensity with viral titer. Pearson correlation coefficient was 0.938 and 0.997, respectively. The cut-off was calculated from the mean of positive control for neutralization plus 3 times the standard deviation, resulting in 0.52% and 14.19, respectively. Before vaccination, individuals presented 60-70% of neutralizing capacity. Thirty days after the first dose, there was a significative increasing of neutralizing capacity in individuals vaccinated with BNT162b2 and ChAdOx1 (*p< 0.0284 and ****p<0.0001, respectively), but no difference with CoronaVac. Thirty days after the second dose, all vaccinated presented a high neutralizing capacity of 85-90%. These percentages comprising an increasing of approximately 20% of nAbs compared to before vaccination (***p<0.0009 for CoronaVac and ****p<0.0001 for BNT162b2 and ChAdOx1). Conclusion:We developed and validated an image-based high content assay, which is safe, highthroughput compatible and able to determine the plasm neutralizing potency in two days.
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