Dengue fever is the most prevalent vector-borne disease in the world, with nearly 100 million people infected every year. Early diagnosis and identification of the pathogen are crucial steps for the treatment and for prevention of the disease, mainly in areas where the co-circulation of different serotypes is common, increasing the outcome of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Due to the lack of fast and inexpensive methods available for the identification of dengue serotypes, herein we report the development of an electrochemical DNA biosensor for the detection of sequences of dengue virus serotype 3 (DENV-3). DENV-3 probe was designed using bioinformatics software and differential pulse voltammetry (DPV) was used for electrochemical analysis. The results showed that a 22-m sequence was the best DNA probe for the identification of DENV-3. The optimum concentration of the DNA probe immobilized onto the electrode surface is 500 nM and a low detection limit of the system (3.09 nM). Moreover, this system allows selective detection of DENV-3 sequences in buffer and human serum solutions. Therefore, the application of DNA biosensors for diagnostics at the molecular level may contribute to future advances in the implementation of specific, effective and rapid detection methods for the diagnosis dengue viruses.
A biosensor that relies on the adsorption immobilization of the 18-mer single-stranded nucleic acid related to dengue virus gene 1 on activated pencil graphite was developed. Hybridization between the probe and its complementary oligonucleotides (the target) was investigated by monitoring guanine oxidation by differential pulse voltammetry (DPV). The pencil graphite electrode was made of ordinary pencil lead (type 4B). The polished surface of the working electrode was activated by applying a potential of 1.8 V for 5 min. Afterward, the dengue oligonucleotides probe was immobilized on the activated electrode by applying 0.5 V to the electrode in 0.5 M acetate buffer (pH 5.0) for 5 min. The hybridization process was carried out by incubating at the annealing temperature of the oligonucleotides. A time of five minutes and concentration of 1 μM were found to be the optimal conditions for probe immobilization. The electrochemical detection of annealing between the DNA probe (TS-1P) immobilized on the modified electrode, and the target (TS-1T) was achieved. The target could be quantified in a range from 1 to 40 nM with good linearity and a detection limit of 0.92 nM. The specificity of the electrochemical biosensor was tested using non-complementary sequences of dengue virus 2 and 3.
Performing repetitions to failure (RF) is a strategy that might acutely reduce neuromuscular performance, as well as increase the rating of perceived exertion (RPE) and the internal training load (ITL) during and after a resistance training (RT) session. Thus, this study aimed to analyze the acute effects of RF or repetitions not to failure (RNF) on countermovement jump (CMJ) performance and the ITL in trained male adults. Eleven men performed two experimental protocols in randomized order (RF vs. RNF). Under the RF condition, participants performed three sets of the leg extension exercise using 100% of the 10RM load and rest intervals of 180-s between sets. Under the RNF condition, participants were submitted to six sets of five repetitions with the same intensity and an 80-s rest interval between sets in the same exercise. The CMJ test was analyzed before and following (15-s and 30-min, respectively) each experimental session. The ITL was evaluated by multiplying the RPE and the total session time, 30-min after the protocol. No main effect or interaction time vs. condition was found for CMJ performance (p > 0.05). In contrast, the ITL showed higher values under the RF condition (p = 0.003). Therefore, even though RF-induced a greater ITL, our results suggest that adopting this strategy in one single-joint exercise for the lower limbs does not seem sufficient to reduce CMJ height.
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