Biomimetic Miniaturized Platform Able to Sustain Arrays of Liquid Droplets for High‐Throughput Combinatorial Tests

The development of high‐throughput and combinatorial technologies is helping to speed up research that is applicable in many areas of chemistry, engineering, and biology. A new model is proposed for flat devices for the high‐throughput screening of accelerated evaluations of multiplexed processes and reactions taking place in aqueous‐based environments. Superhydrophobic (SH) biomimetic surfaces based on the so‐called lotus effect are produced, onto which arrays of micro‐indentations allow the fixing of liquid droplets, based on the rose‐petal effect. The developed platforms are able to sustain arrays of quasi‐spherical microdroplets, allowing the isolation and confinement of different combinations of substances and living cells. Distinct compartmentalized physical, chemical, and biological processes may take place and be monitored in each droplet. The devices permit the addition/removal of liquid and mechanical stirring by adding magnetic microparticles into each droplet. By facing the chip downward, it is possible to produce arrays of cell spheroids developed by gravity in the suspended droplets, with the potential to be used as microtissues in drug screening tests.     

Classification of Brazilian honeys by physical and chemical analytical methods and low field nuclear magnetic resonance (LF H NMR)

This study evaluated the possibility of differentiating the botanical origin of honeys using Low Field Nuclear Magnetic Resonance (LF ¹H NMR). Eighty samples of honey from 8 different botanical sources (eucalyptus, “assa-lipto”, oranges, Barbados cherry, cashew tree, “assa-peixe”, “cipó-uva” and polyfloral) were analyzed. A close correlation (p < 0.01) was established between the LF ¹H NMR analysis and physical and chemical measurements, including water content, water activity, pH and color. Bi-exponential fitting of the transverse relaxation (T₂) data revealed two water populations in all samples, T₂₁ and T₂₂, corresponding to relaxation times of 0.6–1.8 ms and 2.3–5.4 ms respectively. The observed differences in the relaxation times suggest that these were influenced by the differences in botanical origins. Good linear correlations were observed between the T₂ and T₂₁ parameters and the physical and chemical data. This study demonstrated that LF ¹H NMR can be a viable technique for use in classifying honeys by their botanical origin.     

Detection and quantification of Aspergillus westerdijkiae in coffee beans based on selective amplification of beta-tubulin gene by using real-time PCR

Aspergillus westerdijkiae is a new species of fungus that was recently dismembered from Aspergillus ochraceus taxon. Most isolates of A. westerdijkiae are able to produce large amounts of a mycotoxin called ochratoxin A (OA). OA has been found in food and beverages, such as coffee. A. westerdijkiae is very similar to A. ochraceus, and several isolates previously identified as A. ochraceus are now identified as A. westerdijkiae. By using sequences of the beta-tubulin gene, we analyzed several isolates from Brazilian coffee bean samples, previously identified as A. ochraceus, to compare with those of A. westerdijkiae. In fact, most (84%) were identified as A. westerdijkiae. Since this species consistently produces large amounts of OA, we developed a specific primer-pair for detecting and quantifying it in coffee beans by using real-time PCR. The primers Bt2Aw-F 5'TGATACCTTGGCGCTTGTGACG and Bt2Aw-R 5'CGGAAGCCTAAAAAATGAAGAG provided an amplicon of 347 bp in all A. westerdijkiae isolates, and no cross-reaction was observed using DNA from A. ochraceus. The sensitivity of real-time PCR was more than 100 times higher than the cfu technique.     

The effect of fluoride therapies on the morphology of bleached human dental enamel

The aim of this in vitro study was to evaluate qualitatively the surface morphology of enamel bleached with 35% hydrogen peroxide (HP) followed by application of fluoridated agents. Forty intact pre molars were randomly distributed into four groups (n = 10), treated as follows: Group I (control group) remained stored in artificial saliva at 37 °C, Group II - 35% HP; Group III - 35% HP + acidulated fluoride (1.23%) and Group IV - 35% HP + neutral fluoride (2%). The experimental groups received three applications of bleaching gel and after the last application all specimens were polished. This procedure was repeated after 7 and 14 days, and during the intervals of applications, the specimens were stored in artificial saliva at 37°C. Scanning electron microscopy (SEM) analysis showed superficial irregularities and porosities to varying degrees in bleached enamel compared to control group. Sample evaluation was made by attributing scores, and data were statistically analyzed using Kruskal-Wallis and Dunn tests (P < 0.05). SEM qualitative investigation demonstrated that 35% hydrogen peroxide affected human dental enamel morphology, producing porosities, depressions, and superficial irregularities at various degrees. These morphological changes were higher after the application of 1.23% acidulated fluoride gel.     

Self-Assembly Nanoparticles of Natural Bioactive Abietane Diterpenes

Different approaches have been reported to enhance penetration of small drugs through physiological barriers; among them is the self-assembly drug conjugates preparation that shows to be a promising approach to improve activity and penetration, as well as to reduce side effects. In recent years, the use of drug-conjugates, usually obtained by covalent coupling of a drug with biocompatible lipid moieties to form nanoparticles, has gained considerable attention. Natural products isolated from plants have been a successful source of potential drug leads with unique structural diversity. In the present work three molecules derived from natural products were employed as lead molecules for the synthesis of self-assembled nanoparticles. The first molecule is the cytotoxic royleanone 7α-acetoxy-6β-hydroxyroyleanone (Roy, 1) that has been isolated from hairy coleus (Plectranthus hadiensis (Forssk.) Schweinf). ex Sprenger leaves in a large amount. This royleanone, its hemisynthetic derivative 7α-acetoxy-6β-hydroxy-12-benzoyloxyroyleanone (12BzRoy, 2) and 6,7-dehydroroyleanone (DHR, 3), isolated from the essential oil of thicket coleus (P. madagascariensis (Pers.) Benth.) were employed in this study. The royleanones were conjugated with squalene (sq), oleic acid (OA), and/or 1-bromododecane (BD) self-assembly inducers. Roy-OA, DHR-sq, and 12BzRoy-sq conjugates were successfully synthesized and characterized. The cytotoxic effect of DHR-sq was previously assessed on three human cell lines: NCI-H460 (IC₅₀ 74.0 ± 2.2 µM), NCI-H460/R (IC₅₀ 147.3 ± 3.7 µM), and MRC-5 (IC₅₀ 127.3 ± 7.3 µM), and in this work Roy-OA NPs was assayed against Vero-E6 cells at different concentrations (0.05, 0.1, and 0.2 mg/mL). The cytotoxicity of DHR-sq NPs was lower when compared with DHR alone in these cell lines: NCI-H460 (IC₅₀ 10.3 ± 0.5 µM), NCI-H460/R (IC₅₀ 10.6 ± 0.4 µM), and MRC-5 (IC₅₀16.9 ± 0.5 µM). The same results were observed with Roy-OA NPs against Vero-E6 cells as was found to be less cytotoxic than Roy alone in all the concentrations tested. From the obtained DLS results, 12BzRoy-sq assemblies were not in the nano range, although Roy-OA NP assemblies show a promising size (509.33 nm), Pdl (0.249), zeta potential (−46.2 mV), and spherical morphology from SEM. In addition, these NPs had a low release of Roy at physiological pH 7.4 after 24 h. These results suggest the nano assemblies can act as prodrugs for the release of cytotoxic lead molecules.     

Total Synthesis of the Endocannabinoid Uptake Inhibitor Guineensine and SAR Studies

Guineensine ((2E,4E,12E)-13-(benzo[d][1,3]dioxol-5-yl)-N-isobutyltrideca-2,4,12-trienamide) is a plant-derived natural product that inhibits reuptake of the endocannabinoid anandamide with sub-micromolar potency. We have established a highly efficient total synthesis of guineensine, which provided the natural product in only five steps from commercially available 3-nonyn-1-ol in 17 % overall yield, relying on the attachment of the benzodioxolyl moiety to the unsaturated fatty acid chain by means of a Suzuki coupling as the key step. Subsequent SAR studies revealed that replacement of the N-isobutyl group in the natural product by various alkyl, arylalkyl, or aryl groups is generally well tolerated, and derivatives could be identified that are slightly more potent anandamide reuptake inhibitors than guineensine itself. In contrast, modifications of the benzodioxolyl moiety led to decreased activity. Intriguingly, a change in the configuration of the C4=C5 double bond from E to Z was found to be very well tolerated, in spite of the associated change in the overall geometry of the molecule.     

Profile Extrusion: Experimental Assessment of a Numerical Code to Model the Temperature Evolution in the Cooling/Calibration Stage

Designing an industrial calibration system that ensures fast and uniform cooling of a complex extruded profile is always a difficult task. Numerical tools are a valuable aid for the design of these systems in order to achieve fast, low‐cost, and effective solutions. This work comprises the experimental validation of a numerical code, previously developed to support the design of profile extrusion calibration systems, using data collected from an industrial extrusion line. The referred numerical code was developed in the open‐source computational library, OpenFOAM, being able to predict the heat exchanges between the polymer and the calibrator during the profile extrusion cooling stage. The good agreement obtained between the numerical code predictions and the experimental measurements allowed us to confirm the code accuracy. Subsequently, and in order to demonstrate the usefulness of the numerical code, a numerical study was carried out based on an optimization algorithm aimed at improving the performance of the studied calibration system. As a result, the study led to the proposal of an alternative setup, which presents a simpler constructive solution and a better performance than the existing one. POLYM. ENG. SCI., 59:2367–2376, 2019. © 2019 Society of Plastics Engineers     

The effect of water on the performance of TiO2 in photocatalytic selective alkane oxidation

Deactivation of TiO₂ is hampering practical implementation of photocatalytic alternatives for energetically intensive selective oxidation processes. In the present study, humidification of the air stream is demonstrated to be a solution to this problem for well-defined photocatalysts, such as Solaronix TiO₂, in selective oxidation of cyclohexane. Water establishes enhanced product desorption and stimulates decomposition of deactivating surface carboxylate species, as clearly demonstrated by ATR-FTIR spectroscopy. For high surface area TiO₂, such as Hombikat UV100, water molecules adsorb too strongly, and addition of water vapor to the air stream is detrimental to catalyst performance.