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Producing double-stranded RNA (dsRNA) represents a bottleneck for the adoption of RNA interference technology in agriculture, and the main hurdles are related to increases in dsRNA yield, production efficiency, and purity. Therefore, this study aimed to optimize dsRNA production in E. coli HT115 (DE3) using an in vivo system. To this end, we designed a new vector, pCloneVR_2, which resulted in the efficient production of dsRNA in E. coli HT115 (DE3). We performed optimizations in the culture medium and expression inducer in the fermentation of E. coli HT115 (DE3) for the production of dsRNA. Notably, the variable that had the greatest effect on dsRNA yield was cultivation in TB medium, which resulted in a 118% increase in yield. Furthermore, lactose induction (6 g/L) yielded 10 times more than IPTG. Additionally, our optimized up-scaled protocol of the TRIzol™ extraction method was efficient for obtaining high-quality and pure dsRNA. Finally, our optimized protocol achieved an average yield of 53.3 µg/mL after the production and purification of different dsRNAs, reducing production costs by 72%.

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BACKGROUND: The process of tissue injury in coronary artery disease (CAD) has been associated with activation of the complement system, partly due to the action of mannose-binding lectin (MBL) and C3, which are expressed in atherosclerotic lesions. OBJECTIVE: The aim of this study was to evaluate the serum levels of MBL and C3 in patients with CAD and to compare them with healthy controls. Additionally, we aim to assess the correlation between MBL and C3 levels and cardiometabolic parameters. METHODS: MBL and C3 serum concentration were determined by ELISA and immunoturbidimetry, respectively, in up to 119 patients undergoing coronary angiography for CAD evaluation, comprising 48 individuals diagnosed with acute myocardial infarction (MI) and 71 without MI. A total of 93 paired healthy controls were included in the study. RESULTS: Individuals with CAD had MBL serum concentration higher than controls (p = .002), regardless of the presence of MI (p = .006). In addition, high concentration of MBL (>2000 ng/mL) was more frequent in patients with CAD (p = .007; OR = 2.6; 95% CI = 1.3-5.1). C3 levels were not significantly associated with any of the patient groups but were positively correlated with cardiometabolic parameters such as body mass index (BMI) and triglycerides levels. CONCLUSIONS: Higher concentrations of MBL were found to be associated with CAD, whereas C3 levels were found to be associated with cardiovascular risk factors.

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Electrochemotherapy (ECT) and Irreversible electroporation (IRE) are cancer treatments based on electric field distribution in tissues. Solanum tuberosum (potato tissue) phantom is known to mimic changes in the electrical conductivity that occur in animal tissues during electroporation (EP). Electric field distribution is assessed through enzymatic staining. However, the 24-h wait for this assessment could slow agile response scenarios. We developed and validated the Musa acuminata (cavendish banana) conductivity model, which quickly evaluates EP by tissue staining. We investigated the frequency response of the tissue using impedance spectroscopy analysis, conductivity changes, and enzymatic staining. We optimized three usual EP models: adapted Gompertz, smoothed Heaviside, and the sigmoid or logistic function. We found dielectric parameters in banana tissue similar to those in potato (electrical conductivity of 0.035 S/m and relative permittivity of 4.1×104). The coefficients of determination R2 were 99.94% (Gompertz), 99.85% (Heaviside), and 99.58% (sigmoid). The sigmoid and Heaviside functions described the calibration and validation electric currents with 95% confidence. We observed the electroporated areas in bananas 3h30m after EP. Staining was significant after 450 V/cm. The conductivity model of Musa acuminata suits treatment planning, hardware development, and training scenarios. Banana phantom supports the 3Rs practice and is a reliable alternative for potato in EP studies.

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CONTEXT: Many studies show the importance of evaluating the adaptation time of subjects in a virtual driving environment, looking forwards to a response as closest as a possible real vehicle. OBJECTIVES: This study aimed to identify and analyze the adaptation to the driving simulator in older adults and middle-aged adults with and without a distraction, and a secondary aim was to identify predictors of safe performance for older adults' drives. DESIGN: Male and female middle-aged adults (n = 62, age = 30.3 ± 7.1 years) and older adults (n = 102, age = 70.4 ± 5.8 years) were evaluated for braking time performance in a driving simulator; cognition performance assessment included the Mini-Mental State Examination; motor evaluation included ankle flexor muscle strength with the isokinetic dynamometer and handgrip strength; the postural balance was evaluated with Timed Up and Go test, with and without a cognitive distraction task. RESULTS: Older adults (men and women) and middle-aged adult women require more time to adapt to the driving simulator. The distractor increases the adaptation time for all groups. The main predictors of braking time for older women are age, muscle strength, and postural balance associated with distraction, and for older men, muscle strength. CONCLUSIONS: Age, sex, and distractor interfere in the adaptation of the virtual task of driving in a simulator. The evaluation model developed with multi-domains demonstrated the ability to predict which skills are related to braking time with and without the presence of the distractor.

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ABSTRACT Purpose: The aim of this study was to evaluate the fitting process of a scleral lens that allows several parameter adjustments during trials and after the initial period of use. In addition, we verified which adjustments were needed and used the most, their indications, and how often these resources were used, and checked the results. Methods: Scleral contact lens fittings in a private clinic setting were prospectively analyzed in a sequential, non-randomized, and non-comparative manner. All the patients underwent a complete ophthalmic examination and had an indication for scleral lens use (Zenlens, Alden Optical). Results: Scleral fit was analyzed in 80 eyes of 45 patients. Regarding diagnosis, 72% of the patients had keratoconus; 12%, radial keratotomy; 5%, post-refractive surgery ectasia; 5%, dry eye; and 3%, high myopia. In 66 (82.5%) of the 80 eyes studied, parameters were modified when the lenses were ordered. The reasons that led to the modifications were apical touch or decreased sagittal height, increased sagittal height, cylindrical over-refraction, poor visual acuity, lens flexure, peripheral touch, 360° edge compression, horizontal edge compression, and vertical edge compression. Conclusion: In this study, the use of Zenlens scleral lenses was shown to be a promising corrective treatment for patients requiring the use of scleral lenses. Although the study suggests a learning curve, as many adjustments were allowed, the lens could be customized according to the patients' needs. This increased the success rates of fitting and wearing, and consequently, use of the lens became a great option for the visual rehabilitation of patients.

RESUMO Objetivo: Avaliar o processo de adaptação de uma lente escleral que permite vários ajustes de parâmetros durante os testes e após o período inicial do seu uso; verificar quais os ajustes foram necessários, quais foram os mais utilizados, as suas indicações, a frequência com que estes recursos foram utilizados, e avaliar os resultados das mudanças realizadas. Métodos: A adaptação da lente de contato escleral foi analisada prospectivamente, de forma sequencial, não aleatória e não comparativa. Todos os pacientes foram submetidos a um exame oftalmológico completo e tinham indicação para o uso de lentes esclerais. Foi utilizada a lente Zenlens (Alden Optical). Resultados: Foi analisada a adaptação de lentes de contato esclerais em 80 olhos de 45 pacientes. Quanto ao diagnóstico, 72% tinham ceratocone, 12% tinham sido submetidos a ceratotomia radial, 5% tinham ectasia pós-cirurgia refrativa, 5% tinham olho seco, e 3%, alta miopia. Em 66 dos 80 olhos estudados (82,5%), os parâmetros foram modificados quando as lentes foram encomendadas. As razões foram: toque apical ou diminuição da altura sagital, aumento da altura sagital, sobre-refração cilíndrica, baixa acuidade visual, flexão da lente, toque periférico, compressão da borda em 360° e compressão da borda horizontal e/ou vertical. Conclusão: O uso de lentes esclerais Zenlens demonstrou ser uma forma de correção muito promissora para os pacientes que requerem o uso de lentes esclerais. Embora o estudo sugira uma curva de aprendizagem, é possível personalizar as lentes de acordo com as necessidades de cada pacientes. Este fato melhora a adaptação e aumenta a chance de sucesso do uso.

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PURPOSE: The aim of this study was to evaluate the fitting process of a scleral lens that allows several parameter adjustments during trials and after the initial period of use. In addition, we verified which adjustments were needed and used the most, their indications, and how often these resources were used, and checked the results. METHODS: Scleral contact lens fittings in a private clinic setting were prospectively analyzed in a sequential, non-randomized, and non-comparative manner. All the patients underwent a complete ophthalmic examination and had an indication for scleral lens use (Zenlens, Alden Optical). RESULTS: Scleral fit was analyzed in 80 eyes of 45 patients. Regarding diagnosis, 72% of the patients had keratoconus; 12%, radial keratotomy; 5%, post-refractive surgery ectasia; 5%, dry eye; and 3%, high myopia. In 66 (82.5%) of the 80 eyes studied, parameters were modified when the lenses were ordered. The reasons that led to the modifications were apical touch or decreased sagittal height, increased sagittal height, cylindrical over-refraction, poor visual acuity, lens flexure, peripheral touch, 360° edge compression, horizontal edge compression, and vertical edge compression. CONCLUSION: In this study, the use of Zenlens scleral lenses was shown to be a promising corrective treatment for patients requiring the use of scleral lenses. Although the study suggests a learning curve, as many adjustments were allowed, the lens could be customized according to the patients' needs. This increased the success rates of fitting and wearing, and consequently, use of the lens became a great option for the visual rehabilitation of patients.

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Abstract Context: Many studies show the importance of evaluating the adaptation time of subjects in a virtual driving environment, looking forwards to a response as closest as a possible real vehicle. Objectives: This study aimed to identify and analyze the adaptation to the driving simulator in older adults and middle-aged adults with and without a distraction, and a secondary aim was to identify predictors of safe performance for older adults' drives. Design: Male and female middle-aged adults (n = 62, age = 30.3 ± 7.1 years) and older adults (n = 102, age = 70.4 ± 5.8 years) were evaluated for braking time performance in a driving simulator; cognition performance assessment included the Mini-Mental State Examination; motor evaluation included ankle flexor muscle strength with the isokinetic dynamometer and handgrip strength; the postural balance was evaluated with Timed Up and Go test, with and without a cognitive distraction task. Results: Older adults (men and women) and middle-aged adult women require more time to adapt to the driving simulator. The distractor increases the adaptation time for all groups. The main predictors of braking time for older women are age, muscle strength, and postural balance associated with distraction, and for older men, muscle strength. Conclusions: Age, sex, and distractor interfere in the adaptation of the virtual task of driving in a simulator. The evaluation model developed with multi-domains demonstrated the ability to predict which skills are related to braking time with and without the presence of the distractor.

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OBJECTIVE: To assess respiratory system impedance (Zrs) and spirometric parameters in children and adolescents with and without a history of preterm birth. METHODS: We evaluated a sample of 51 subjects between 11 and 14 years of age: 35 who had a history of preterm birth (preterm group) and 16 who had been born at term (full-term group). Lung function was measured by spirometry, spectral oscillometry, and intra-breath oscillometry. RESULTS: Neither spirometry nor spectral oscillometry revealed any statistically significant differences between the preterm and full-term groups. However, intra-breath oscillometry demonstrated significant differences between the two groups in terms of the change in resistance, reactance at end-inspiration, and the change in reactance (p < 0.05 for all). CONCLUSIONS: Our findings suggest that abnormalities in Zrs persist in children and adolescents with a history of preterm birth and that intra-breath oscillometry is more sensitive than is spectral oscillometry. Larger studies are needed in order to validate these findings and to explore the impact that birth weight and gestational age at birth have on Zrs later in life.

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The rotary jet spinning technique permits the production of biomaterials that can be used as devices that come into contact with biological systems (including biological fluids) for diagnostic or surgical applications. These materials are composed of synthetic or natural compounds and allow the incorporation of drugs for therapeutic purposes. Two solutions containing 50% poly(lactic acid) (PLA) and 50% poly(ε-caprolactone) (PCL) diluted in three different solvents were prepared for rotary jet spinning (RJS) process. Vancomycin, an antibiotic indicated for the treatment of severe staphylococcal infections in patients with penicillin allergy, was added in the polymer solutions, to obtain drug-loaded fibrous mats. Morphological surface characterization by scanning electron microscopy revealed heterogeneous pores in the microfibers. Vancomycin loading interfered with the morphology of all samples in terms of fiber size, leading to smaller diameter fibers. Attenuated total reflectance/Fourier transform infrared spectroscopy was used for identification of the samples. The vibrational characteristics of PCL/PLA and vancomycin were consistent with expectations. Vero cell culture assays by the extract dilution and direct contact methods revealed the absence of cytotoxicity, except for the sample prepared with 50% of PCL and of a 9/2 (V/V) vancomycin content, with the growth of confluent and evenly spread cells on the fibrous mats surface. Microbiological analysis, performed on Staphylococcus aureus by the halo inhibition test and by the broth dilution method, showed that the antibacterial activity of vancomycin was maintained by the loading process in the polymer fibers. The results showed that rotary jet spinning produces satisfactory amounts of Vancomycin-loaded fibers, as potential web dressing for wound repair.

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ABSTRACT Objective: To assess respiratory system impedance (Zrs) and spirometric parameters in children and adolescents with and without a history of preterm birth. Methods: We evaluated a sample of 51 subjects between 11 and 14 years of age: 35 who had a history of preterm birth (preterm group) and 16 who had been born at term (full-term group). Lung function was measured by spirometry, spectral oscillometry, and intra-breath oscillometry. Results: Neither spirometry nor spectral oscillometry revealed any statistically significant differences between the preterm and full-term groups. However, intra-breath oscillometry demonstrated significant differences between the two groups in terms of the change in resistance, reactance at end-inspiration, and the change in reactance (p < 0.05 for all). Conclusions: Our findings suggest that abnormalities in Zrs persist in children and adolescents with a history of preterm birth and that intra-breath oscillometry is more sensitive than is spectral oscillometry. Larger studies are needed in order to validate these findings and to explore the impact that birth weight and gestational age at birth have on Zrs later in life.

RESUMO Objetivo: Avaliar a impedância do sistema respiratório (Zsr) e parâmetros espirométricos em crianças e adolescentes com e sem história de prematuridade. Métodos: Foi analisada uma amostra de 51 indivíduos entre 11 e 14 anos de idade: 35 com história de prematuridade (grupo pré-termo) e 16 nascidos a termo (grupo a termo). A função pulmonar foi medida por meio de espirometria, oscilometria espectral e oscilometria intra-breath. Resultados: A espirometria e a oscilometria espectral não revelaram diferenças estatisticamente significativas entre os grupos pré-termo e a termo. No entanto, a oscilometria intra-breath demonstrou diferenças significativas entre os dois grupos quanto à alteração da resistência, à reatância ao final da inspiração e à alteração da reatância (p < 0,05 para todas). Conclusões: Nossos achados sugerem que as anormalidades na Zsr persistem em crianças e adolescentes com história de prematuridade e que a oscilometria intra-breath é mais sensível do que a oscilometria espectral. São necessários estudos maiores para validar esses achados e para explorar o impacto do peso e idade gestacional ao nascer na Zsr mais tarde na vida.