ß-Cyclodextrins as affordable antivirals to treat coronavirus infection.

The SARS-CoV-2 pandemic made evident that there are only a few drugs against coronavirus. Here we aimed to identify a cost-effective antiviral with broad spectrum activity and high safety profile. Starting from a list of 116 drug candidates, we used molecular modelling tools to rank the 44 most promising inhibitors. Next, we tested their efficacy as antivirals against α and ß coronaviruses, such as the HCoV-229E and SARS-CoV-2 variants. Four drugs, OSW-1, U18666A, hydroxypropyl-ß-cyclodextrin (HßCD) and phytol, showed in vitro antiviral activity against HCoV-229E and SARS-CoV-2. The mechanism of action of these compounds was studied by transmission electron microscopy and by fusion assays measuring SARS-CoV-2 pseudoviral entry into target cells. Entry was inhibited by HßCD and U18666A, yet only HßCD inhibited SARS-CoV-2 replication in the pulmonary Calu-3 cells. Compared to the other cyclodextrins, ß-cyclodextrins were the most potent inhibitors, which interfered with viral fusion via cholesterol depletion. ß-cyclodextrins also prevented infection in a human nasal epithelium model ex vivo and had a prophylactic effect in the nasal epithelium of hamsters in vivo. All accumulated data point to ß-cyclodextrins as promising broad-spectrum antivirals against different SARS-CoV-2 variants and distant alphacoronaviruses. Given the wide use of ß-cyclodextrins for drug encapsulation and their high safety profile in humans, our results support their clinical testing as prophylactic antivirals.

Impact of Non-Thermal Technologies on the Quality of Nuts: A Review.

Nuts are widely consumed worldwide, mainly due to their characteristic flavor and texture, ease of consumption, and their functional properties. In addition, consumers increasingly demand natural or slightly processed foods with high quality. Consequently, non-thermal treatments are a viable alternative to thermal treatments used to guarantee safety and long shelf life, which produce undesirable changes that affect the sensory quality of nuts. Non-thermal treatments can achieve results similar to those of the traditional (thermal) ones in terms of food safety, while ensuring minimal loss of bioactive compounds and sensory properties, thus obtaining a product as similar as possible to the fresh one. This article focuses on a review of the main non-thermal treatments currently available for nuts (cold plasma, high pressure, irradiation, pulsed electric field, pulsed light, ultrasound and ultraviolet light) in relation to their effects on the quality and safety of nuts. All the treatments studied have shown promise with regard to the inhibition of the main microorganisms affecting nuts (e.g., Aspergillus, Salmonella, and E. coli). Furthermore, by optimizing the treatment, it is possible to maintain the organoleptic and functional properties of these products.

Effect of pH on pulsed light inactivation of polyphenol oxidase.

The inactivation of diverse food enzymes by pulsed light (PL) has been described before, including the inactivation of polyphenol oxidase (PPO) (at pH 6.5). Since the pH affects the conformation of enzymes, it may influence the inactivation of enzymes by PL. The aim of this work was to evaluate the effect of pH on the kinetics of the PL-inactivation and associated structural changes of a case enzyme. To this, PPO was treated by PL at different pHs (4.0-6.5) and its inactivation kinetics and changes in its structure were evaluated by spectrophotometric and spectrofluorometric methods. The inactivation proceeded faster at low pH and was highly correlated with the decrease in peak intrinsic fluorescence intensity. Phase diagrams and parameter A evolution indicated the absence of intermediate unfolded states during the course of the inactivation. No protein aggregation was detected by turbidimetry. Results indicate that although a low pH favors the PL-inactivation of PPO, the mechanism of inactivation is pH-independent. Beyond the specific outcome for PPO, the results are evidence of a general pH-independence in the mechanism of enzyme inactivation by PL in the pH range 4.0-6.5 and acidification can be a strategy to decrease treatment times during PL processing.

Evaluation of Pulsed Light to Inactivate Brettanomyces bruxellensis in White Wine and Assessment of Its Effects on Color and Aromatic Profile.

Brettanomyces bruxellensis is a wine spoilage yeast that could be inactivated by pulsed light (PL); however, this technology may induce changes in the quality of this alcoholic drink. The present research aimed to determine the potential of PL to inactivate B. bruxellensis inoculated in white wine and to assess the effect of this technology on the color and aromatic profile of the wine. For this, a cocktail of B. bruxellensis strains was inoculated into the wine and its inactivation by PL was determined and fitted to a microbial inactivation model. Along with this, the effect of PL on instrument-measured color, and the volatile compounds of the wine were evaluated by GC/MS and descriptive sensory analysis, respectively. B. bruxellensis was inactivated according to the Geeraerd model including the tail effect, with a maximum inactivation of 2.10 log reduction at 10.7 J/cm2; this fluence was selected for further studies. PL affected wine color but the total color difference was below the just noticeable difference at 10.7 J/cm2. The concentration of 13 out of 15 volatile compounds decreased due to the PL, which was noticeable by the panel. It is not clear if these compounds were photolyzed or volatilized in the open reactor during treatment. In conclusion, PL is able to inactivate B. bruxellensis in white wine but the treatment impairs the volatile profile. The use of a closed reactor under turbulent flow is recommended for disaggregating yeast clumps that may cause the tailing of the inactivation curve, and to avoid the possible escape of volatile compounds during treatment.

Synthesis of New Cyclodextrin-Based Adsorbents to Remove Direct Red 83:1.

Two cyclodextrins (CDs), γ- and hydroxypropyl (HP)-γ-CDs were used to synthesize new adsorbents by using epichlorohydrin (EPI) as cross-linking agent in order to remove Direct Red 83:1 (DR) from water. Both polymers were characterized in terms of Fourier spectroscopy, nuclear magnetic resonance, particle size distribution and thermogravimetric analysis. Experimental data for both polymers were well fitted to the pseudo-second order and intraparticle diffusion model, indicating that in the adsorption both chemical and physical interactions are essential in the removal of DR. Three different isotherm models were analyzed, concluding that γ-CDs-EPI followed the Temkin isotherm and HP-γ-CDs-EPI the Freundlich isotherm, these results suggested that the adsorption was happening onto heterogeneous surfaces. The results of the Gibbs free energy showed that the adsorption was spontaneous at room temperature. In order to eliminate the remaining dye after the polymer treatment, and advanced oxidation process (AOP) was considered, achieving more than 90% of removal combining both mechanisms.

Egg By-Products as a Tool to Remove Direct Blue 78 Dye from Wastewater: Kinetic, Equilibrium Modeling, Thermodynamics and Desorption Properties.

Eggshell, a waste material from food manufacturing, can be used as a potential ecofriendly adsorbent for the elimination of textile dyes from water solutions. The adsorption process was evaluated varying factors such as initial dye load, contact time, pH, quantity of adsorbent, and temperature. The initial dye load (Direct Blue 78) was in the range of 25-300 mg/L. The kinetics of adsorption were analyzed using different models, such as pseudo-first-order, pseudo-second-order, and intraparticle diffusion model. Also, the experimental data at equilibrium were studied using Freundlich, Langmuir, and Temkin isotherms. The kinetics followed pseudo-second-order, then pseudo-first-order, and finally the model of intraparticle diffusion. The results obtained for data at equilibrium follow the order: Freundlich > Langmuir > Temkin. The adsorption equilibrium showed a maximum capacity of adsorption (qmax) of 13 mg/g at pH 5, and using 0.5 g of eggshell. Dye adsorption was enhanced with increasing temperatures. The thermodynamic study revealed the spontaneity and endothermic nature of the adsorption process. The desorption study shows that the eggshell could be reused in different adsorption/desorption cycles. A novel advanced oxidation process could degrade more than 95% of the dye. The results show that eggshell is a waste material useful to remove hazardous dyes from wastewater, which may alleviate the environmental impact of dyeing industries.

Adsorption of Direct Blue 78 Using Chitosan and Cyclodextrins as Adsorbents.

The dyeing industry is one of the most polluting in the world. The adsorption of dyes by polymeric matrixes can be used to minimize the discharge of dyes into the environment. In the present study, chitosan-NaOH and ß-cyclodextrin-epichlorohydrin polymers were used to remove the dye Direct Blue 78 from a wastewater model. To understand the adsorption behavior of Direct Blue 78 onto the polymers, adsorption rate and maximum adsorption capacity were calculated using kinetic tests and isotherm curves respectively. The kinetic data and mechanism of the adsorption process were analyzed by three models and the equilibrium data by three adsorption isotherms; also the different thermodynamic parameters were calculated. Results showed that the adsorption process follows pseudo-second-order kinetics in both polymers and the Langmuir isotherm best-fitted data for chitosan-NaOH polymer and the Freundlich isotherm for the ß-CDs-EPI polymer. The adsorption process is exothermic in both cases and spontaneous for the ß-CDs-EPI polymer to a certain temperature and not spontaneous for the chitosan-NaOH polymer and ß-CDs-EPI polymer at higher temperatures. The complementary action of an advanced oxidation process eliminated >99% of the dye from water. The coupled process seems to be suitable for reducing the environmental impact of the dyeing industry.

Adsorption Properties of ß- and Hydroxypropyl-ß-Cyclodextrins Cross-Linked with Epichlorohydrin in Aqueous Solution. A Sustainable Recycling Strategy in Textile Dyeing Process.

ß-cyclodextrin (ß-CD) and hydroxypropyl-ß-cyclodextrin (HP-ß-CD) were used to prepare insoluble polymers using epichlorohydrin as a cross-linking agent and the azo dye Direct Red 83:1 was used as target adsorbate. The preliminary study related to adsorbent dosage, pH, agitation or dye concentration allowed us to select the best conditions to carry out the rest of experiments. The kinetics was evaluated by Elovich, pseudo first order, pseudo second order, and intra-particle diffusion models. The results indicated that the pseudo second order model presented the best fit to the experimental data, indicating that chemisorption is controlling the process. The results were also evaluated by Freundlich, Langmuir and Temkin isotherms. According to the determination coefficient (R²), Freunlich gave the best results, which indicates that the adsorption process is happening on heterogeneous surfaces. One interesting parameter obtained from Langmuir isotherm is qmax (maximum adsorption capacity). This value was six times higher when a ß-CDs-EPI polymer was employed. The cross-linked polymers were fully characterized by nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TGA). Also, morphology and particle size distribution were both assessed. Under optimized conditions, the ß-CDs-EPI polymer seems to be a useful device for removing Direct Red 83:1 (close 90%), from aqueous solutions and industrial effluents. Complementarily, non-adsorbed dye was photolyzed by a pulsed light driven advanced oxidation process. The proposed methodology is environmental and economically advantageous, considering the point of view of a sustainable recycling economy in the textile dyeing process.

Thorough characterization and stability of HP-ß-cyclodextrin thymol inclusion complexes prepared by microwave technology: A required approach to a successful application in food industry.

BACKGROUND: The aims of the present study were to obtain a stable dry powder formulation of cyclodextrins (CDs) encapsulating thymol, for successful use as an ingredient on an industrial scale, and to characterize the thymol-CDs complexes using different techniques. RESULTS: Thymol was successfully solubilized in aqueous solutions and the Kc value increased with the pH of the media until the pH was neutral, giving the highest values (2583 ± 176 L mol-1 ) for HP-ß-cyclodextrins (HP-ß-CDs). The best encapsulation efficiency of thymol in solid complexes was obtained using the microwave (MWI) encapsulation method. The different characterization techniques have demonstrated the affinity of HP-ß-CDs for thymol molecules, forming stable complexes. CONCLUSIONS: The results support the use of the MWI method in the preparation of solid HP-ß-CD-thymol complexes, due to greater encapsulation efficiency and technological and economic advantages for industrial applications. © 2018 Society of Chemical Industry.

Microbiological and Sensory Quality of Fresh Ready-to-Eat Artichoke Hearts Packaged under Modified Atmosphere.

In recent years the sales of minimally processed vegetables have grown exponentially as a result of changes in consumer habits. The availability of artichoke buds as a ready-to-eat product would be, therefore, highly advantageous. However, minimally processed artichoke hearts are difficult to preserve because of their rapid browning and the proliferation of naturally occurring microorganisms. We developed artichoke hearts prepared as ready-to-eat products that maintain the characteristics of the fresh product. The microbiological stability, sensory qualities, and shelf life of the processed artichoke hearts were determined. During the shelf life, Salmonella, Listeria monocytogenes, and Escherichia coli counts were below the limits legally established by European regulations for minimally processed vegetables. The pH played an important role in microbial growth. Artichoke hearts had lower microbial counts in experiments conducted at pH 4.1 than in experiments conducted at pH 4.4, although the recommended threshold value for total plate count (7 log CFU/g) was not exceeded in either case. Sensory parameters were affected by the microorganisms, and artichoke products at lower pH had better sensory qualities. Vacuum impregnation techniques, modified atmosphere packaging, and low storage temperature were very effective for increasing the shelf life of minimally processed artichokes. The average shelf life was approximately 12 to 15 days.

Detection of mandarin in orange juice by single-nucleotide polymorphism qPCR assay.

A dual-probe real time PCR (qPCR) DNA-based analysis was devised for the identification of mandarin in orange juice. A single nucleotide polymorphism at the trnL-trnF intergenic region of the chloroplast chromosome was confirmed in nine orange (Citrus sinensis) and thirteen commercial varieties of mandarin, including Citrus reticulata and Citrus unshiu species and a mandarin × tangelo hybrid. Two short minor-groove binding fluorescent probes targeting the polymorphic sequence were used in the dual-probe qPCR, which allowed the detection of both species in single-tube reactions. The similarity of PCR efficiencies allowed a simple estimation of the ratio mandarin/orange in the juice samples, which correlated to the measured difference of threshold cycle values for both probes. The limit of detection of the assay was 5% of mandarin in orange juice, both when the juice was freshly prepared (not from concentrate) or reconstituted from concentrate, which would allow the detection of fraudulently added mandarin juice. The possible use of the dual-probe system for quantitative measurements was also tested on fruit juice mixtures. qPCR data obtained from samples containing equal amounts of mandarin and orange juice revealed that the mandarin target copy number was approximately 2.6-fold higher than in orange juice. The use of a matrix-adapted control as calibrator to compensate the resulting C(T) bias allowed accurate quantitative measurements to be obtained.

Arrays on disc for screening and quantification of pollutants.

A rapid compact disc based methodology for screening and quantification of organic pollutants in mandarin juices is presented. The assay is established on the coating conjugate indirect competitive principle and developed under disc-array configuration. The detection is based on the acquisition of attenuated reflective signals that were proportional to optical density of the immunoreaction product. The competitive assay is applied to quantify simultaneously, in a selective manner, non-systemic insecticides in mandarin juices. The detection limits were 0.2 and 0.1 µg L(-1) and the sensitivity 2.1 and 1.5 µg L(-1), for chlorpyrifos and fenthion, respectively. Pollutants were directly quantified after sample dilution in a total time of 40 min. Also, the implementation of positive and negative controls into the array configuration served as an automatic quality control test. The effect of thermal treatment on pesticide dissipation was studied and found that it was insignificant under the studied conditions. Recovery intervals ranged from 96-105% to 94-103%, for chlorpyrifos and fenthion, respectively and were similar to those obtained with gas chromatography coupled to mass spectrometry. In the current configuration, 64 samples can be simultaneously analyzed on a disc at a very competitive value, demonstrating its potential for high-throughput multiplexed screening applications for controlled monitoring programs in low-level labs or outside the lab setting.

Effects of cyclodextrin type on vitamin C, antioxidant activity, and sensory attributes of a mandarin juice enriched with pomegranate and goji berries.

UNLABELLED: The effects of the addition of cyclodextrins (CDs), ß-CD, or HP-ß-CD (1%), on the protection of antioxidant compounds of mandarin juices enriched with pomegranate extract and goji berries juice, was studied. Juices were prepared and after their thermal treatment (98 °C, 30 s) they were stored at 4 °C during 75 d. Vitamin C content, CIE L*a*b* color, antioxidant capacity, retinol equivalents, and sensory properties were studied. Losses on vitamin C were higher (6%) for juices with ß-CD than juices with HP-ß-CD. Retinol equivalents degradation was lower (3.4%) in juices with HP-ß-cyclodextrins than in those treated with ß-CD. Lower losses were observed for the instrumental and sensory color intensity in juices with HP-ß-CD addition. Finally, the antioxidant capacity was also higher in juices treated with HP-ß-CD. Finally, the overall sensory quality of juices with HP-ß-CD was the best one after 30 d of cold storage. Even though ß-CD addition did not cause any improvement compared with control juice (without CD addition), the benefits of adding HP-ß-CD to this particular juice were shown in almost all parameters under study. PRACTICAL APPLICATION: The present study deals with the practical aspects of the utilization of CDs and CD-complexes in the food industry. The molecular encapsulation of lipophilic food ingredients with CD are supposed to improve the stability of flavors, vitamins, colorants, and antioxidant activity, leading to extended product shelf life. The HP-ß-CD treated mandarin juice enriched with pomegranate extract and juice of goji berries will have a more intense color, higher vitamin C content, retinol equivalents, and antioxidant activity during storage and shelf life than control juices, with no CD addition.

Ochratoxin A in red paprika: relationship with the origin of the raw material.

The occurrence of ochratoxin A (OA) in paprika elaborated from peppers grown in several countries (Peru, Brazil, Zimbabwe and Spain) was studied, using an immunoaffinity clean-up column coupled to liquid chromatography and fluorescence detection. The preparation of the methyl ester (OA-Me) and liquid chromatography-electrospray-ion trap-mass spectrometry was used both to confirm the identity of the chromatographic peak that correspond to OA and to quantify it at low levels or in dirty fractions. A total of 115 strains of moulds were isolated; 85 of the fungal strains were obtained from OA contaminated paprika samples and identified as belonging to the Aspergillus Section Circumdati group (A. ochraceus) and Section Nigri group (A. niger, A. carbonarius). Among the latter ones, 31% of the A. ochraceus isolates and one A. niger were OA producers in vitro. None of the mould strains isolated from paprika samples with undetectable levels of OA or concentrations below 1 microg kg(-1) were toxin producers. Great differences in OA content in paprika samples were found, and a relationship with the climatic conditions of the geographic origin of the samples, and with cultural and technical practices in pepper manipulation is suggested.

Evaluation of a novel malathion immunoassay for groundwater and surface water analysis.

Malathion is an organophosphorus insecticide commonly used in crops and indoor applications. Negative effects of malathion on human health and ecosystems are of growing concern. In this work, novel malathion haptens are synthesized to develop an ELISA screening method. The immunoassay is based on a conjugate-coated format and shows a limit of detection of 0.11 ng/mL, an IC50 of 1.58 ng/mL, a dynamic range between 0.23 and 10.94 ng/ mL, and a cross-reactivity of <2% with structurally related compounds. The developed ELISA has been used to quantify malathion in groundwater and surface water samples. The good recoveries achieved and the agreement with those given by the GC-MS reference method indicate the potential of the assay for environmental monitoring of malathion in natural waters without purification or preconcentration steps. The effect of dissolved organic matter (humic acids) on the ELISA was evaluated, resulting in the conclusion that the immunoassay can be successfully performed in surface water samples with a humic acid content up to 10 mg/L, without sample pretreatment. Samples with a high humic acid content can be analyzed through ELISA after solid-phase extraction, eluting with acetone or acetonitrile.