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According to their origin, major postharvest losses of citrus fruit are caused by weight loss, fungal diseases, physiological disorders, and quarantine pests. Cold storage and postharvest treatments with conventional chemical fungicides, synthetic waxes, or combinations of them are commonly used to minimize postharvest losses. However, the repeated application of these treatments has led to important problems such as health and environmental issues associated with fungicide residues or waxes containing ammoniacal compounds, or the proliferation of resistant pathogenic fungal strains. There is, therefore, an increasing need to find non-polluting alternatives to be used as part of integrated disease management (IDM) programs for preservation of fresh citrus fruit. Among them, the development of novel natural edible films and coatings with antimicrobial properties is a technological challenge for the industry and a very active research field worldwide. Chitosan and other edible coatings formulated by adding antifungal agents to composite emulsions based on polysaccharides or proteins and lipids are reviewed in this article. The most important antifungal ingredients are selected for their ability to control major citrus postharvest diseases like green and blue molds, caused by Penicillium digitatum and Penicillium italicum, respectively, and include low-toxicity or natural chemicals such as food additives, generally recognized as safe (GRAS) compounds, plant extracts, or essential oils, and biological control agents such as some antagonistic strains of yeasts or bacteria.

Starch–gellan (80:20) coating formulations were applied to apples and persimmons to analyse their effectiveness controlling the weight loss, respiration rate, fruit firmness, and fungal decay during postharvest. Thyme essential oil (EO) was incorporated (0.25 and 0.5 g per g of polymer) directly or encapsulated in lecithin to enhance antifungal action. Coatings did not reduce weight loss or firmness changes in apples, but they prevented water loss in persimmons. In contrast, no significant effect of the coatings was observed on the respiration rates and the respiration quotient of persimmons, whereas the respiration rates and quotient was increased in apples. On the other hand, the coatings without lecithin reduced the incidence and severity of black spot caused by Alternaria alternata in persimmons, regardless of the content of essential oil. Likewise, these reduced the severity of gray mold caused by Botrytis cinerea in apples. No positive effect of lecithin in coatings was observed on the postharvest quality and decay in either apples or persimmons, nor did EO exert antifungal action despite its proven effectiveness in in vitro tests.

The curative antifungal activity of edible composite coatings (ECs) based on pregelatinized potato starch-glyceryl monostearate (PPS-GMS) formulated with or without sodium benzoate (SB) to control green mould (caused by Penicillium digitatum) and blue mould (P. italicum) was assessed on 'Orri' mandarins, 'Valencia' oranges and 'Fino' lemons. These fruit were artificially inoculated with P. digitatum or P. italicum, treated by immersion in coating emulsions and compared to uncoated control fruit immersed in water and fruit immersed in 2% SB (w/v) aqueous solution. Treated fruit were then stored at either 20°C or commercial low temperature (5°C for mandarins and oranges, 12°C for lemons). Coatings without SB did not exhibit antifungal activity, whereas coatings containing 2% SB reduced incidence and severity of green and blue moulds, in comparison to the controls, on all citrus species and in all storage conditions, without differing from the aplication of 2% SB alone. For example, incidence reduction on 'Fino' lemons was from 99 to 0% after 7 d at 20°C, and from 99 to 30% after 2 weeks at 12°C. None of the treatments was phytotoxic. These results indicate that applications of SB as antifungal ingredient of PPS-GMS based ECs is a promising non-polluting alternative to control Penicillium postharvest decay of citrus, and these ECs are effective substitutes for conventional waxes amended with synthetic fungicides.

The influence of hydroxypropyl methylcellulose- (HPMC-) beeswax (BW) composite edible coatings formulated with or without food additives with antifungal properties on physicochemical and sensory properties of plums (Prunus salicina) cv. “Friar” stored for 11 and 22 d at 1°C followed by a shelf life period of 5 d at 20°C was evaluated. Food preservatives selected from previous research included potassium sorbate (PS), sodium methyl paraben (SMP), and sodium ethyl paraben (SEP). Emulsions had 7% of total solid content and were prepared with glycerol and stearic acid as plasticizer and emulsifier, respectively. All the coatings reduced plum weight and firmness loss and coated fruit showed higher titratable acidity, soluble solids content, and hue angle values at the end of the storage period. In addition, physiological disorders such as flesh browning and bleeding were reduced in coated samples compared to uncoated controls. Paraben-based coatings were the most effective in controlling weight loss and the SMP-based coating was the most effective in maintaining plum firmness. Respiration rate, sensory flavor, off-flavors, and fruit appearance were not adversely affected by the application of antifungal coatings. Overall, these results demonstrated the potential of selected edible coatings containing antifungal food additives to extend the postharvest life of plums, although further studies should focus on improving some properties of the coatings to enhance gas barrier properties and further increase storability.

To assess the possible interactions between the dorsolateral periaqueductal gray matter (dlPAG) and the different domains of the nucleus ambiguus (nA), we have examined the pattern of double-staining c-Fos/FoxP2 protein immunoreactivity (c-Fos-ir/FoxP2-ir) and tyrosine hydroxylase (TH) throughout the rostrocaudal extent of nA in spontaneously breathing anaesthetised male Sprague–Dawley rats during dlPAG electrical stimulation. Activation of the dlPAG elicited a selective increase in c-Fos-ir with an ipsilateral predominance in the somatas of the loose (p < 0.05) and compact formation (p < 0.01) within the nA and confirmed the expression of FoxP2 bilaterally in all the domains within the nA. A second group of experiments was made to examine the importance of the dlPAG in modulating the laryngeal response evoked after electrical or chemical (glutamate) dlPAG stimulations. Both electrical and chemical stimulations evoked a significant decrease in laryngeal resistance (subglottal pressure) (p < 0.001) accompanied with an increase in respiratory rate together with a pressor and tachycardic response. The results of our study contribute to new data on the role of the mesencephalic neuronal circuits in the control mechanisms of subglottic pressure and laryngeal activity.

The sulfur-containing salts, classified as food additives, sodium metabisulfite (SMBS), potassium metabisulfite (PMBS), aluminum sulfate (AlS), and aluminum potassium sulfate (AlPS), were evaluated for their activity against Monilinia fructicola, Rhizopus stolonifer, and Geotrichum candidum, the most economically important fungal pathogens causing postharvest disease of stone fruit. In in vitro tests with potato dextrose agar (PDA) Petri dishes amended with different concentrations of the salts (0, 10, 20, 30, 50, and 100 mM), SMBS and PMBS at all concentrations, AlS above 20 mM, and AlPS above 30 mM, completely inhibited the mycelial growth of the three fungi after incubation at 25º C for up to 10 days. In in vivo primary screenings with artificially inoculated nectarines, aqueous solutions of the four salts reduced the incidence and severity of brown rot (BR) at concentrations of 10 and 50 mM, whereas only AlS and AlPS reduced Rhizopus rot (RR), and none of the salts was effective against sour rot (SR). Solutions at 100 mM were phytotoxic and injured the fruit peel. In small-scale trials, 1 min dip treatments at 20º C in SMBS or PMBS at 10 mM significantly reduced the incidence and severity of BR after incubation at 20º C for up to 8 days. Conversely, dips in AlS and AlPS reduced neither BR nor RR. Results highlight the potential of SMBS and PMBS as new nonpolluting tools for the integrated control of BR, but not RR and SR, on stone fruit.

The in vitro antifungal activity of various generally recognized as safe (GRAS) salts against Colletotrichum gloeosporioides, the causal agent of citrus postharvest anthracnose, was evaluated as mycelial growth reduction on potato dextrose agar (PDA) dishes amended with salt aqueous solutions at different concentrations. The most effective treatments [0.2% ammonium carbonate (AC), 2% potassium sorbate (PS), 0.2% potassium carbonate (PC), 0.1% sodium methylparaben (SMP), 0.1% sodium ethylparaben (SEP), 2% sodium benzoate (SB) and 2% potassium silicate (PSi)] were selected as antifungal ingredients of composite edible coatings formulated with hydroxypropyl methylcellulose (HPMC)-beeswax (BW) matrixes. Stable coatings containing these salts were applied in in vivo curative experiments to “Nadorcott” mandarins and “Valencia” oranges artificially inoculated with C. gloeosporioides and those containing 2% PS, 2% SB and 2% PSi were the most effective to reduce anthracnose severity with respect to control fruit (up to 70% on mandarins). The effect of these selected coatings on the quality of non-inoculated and cold-stored “Valencia” oranges was determined after 28 and 56 days at 5 ºC and 90% RH, followed by 7 days of shelf life at 20 ºC. None of the coatings significantly reduced weight loss of coated oranges, but they modified their internal atmosphere, increasing the CO2 content. Overall, the coatings did not adversely affect the physicochemical and sensory attributes of the fruit.

Composite edible coatings based on hydroxypropyl methylcellulose (HPMC), as a polymeric phase, and oleic acid (OA) or beeswax (BW), as a hydrophobic phase, were formulated with different food additives as antifungal ingredients. HPMC–OA coatings containing 2% (w/v) sodium benzoate (SB), 1% ammonium carbonate (AC), 1% potassium carbonate (PC), 1% potassium bicarbonate (PBC), 1% sodium bicarbonate (SBC), 1% potassium silicate (PSi), 0.1% sodium methyl paraben (SMP) or 0.1% sodium ethyl paraben (SEP), and HPMC–BW coatings containing 2% sodium propionate (SP), 2% PBC, 2% SB or 0.1% SEP were evaluated for the control of Alternaria black spot (ABS) on Diospyros kaki Thunb. ’Rojo Brillante’ persimmons artificially inoculated with Alternaria alternata. After 14 days of incubation at 20 °C, HPMC–OA coatings formulated with PBC, PC or SEP were the most effective to reduce ABS incidence (61, 54, and 36% reduction, respectively, concerning uncoated control fruit) and severity (28, 12 and 22% reduction, respectively), while only HPMC–BW coatings formulated with SEP significantly reduced ABS incidence (50% reduction) and severity (36% reduction). HPMC–OA and HPMC–BW coatings containing 2% PBC or 0.1% SEP were selected to evaluate their effect on the weight loss, firmness and respiration rate of healthy ‘Rojo Brillante’ persimmons cold-stored at 1 °C and 90% relative humidity (RH) for 15 and 30 days, followed by 7 days of shelf life at 20 °C. HPMC–BW coatings were more effective in reducing fruit weight and firmness losses than HPMC–OA coatings, while all antifungal coatings significantly reduced fruit respiration. Overall, the HPMC–BW edible coating that contains SEP could be a promising postharvest treatment to control ABS and maintain the quality of cold-stored ‘Rojo Brillante’ persimmons.

Stimulation of the dorsolateral periaqueductal grey matter (dlPAG) in rats evokes an active defensive behaviour together with a cardiorespiratory response characterised by tachypnoea, tachycardia and hypertension. The dlPAG neurons involved in these responses are excitatory, presumably glutamatergic, due to the presence of vesicular glutamate transporter VGLUT2 within their axon terminals. Previously, our group described a functional interaction between dlPAG and the pontine A5 region. Accordingly, in the present work, in order to characterize the role of glutamate within this interaction, experiments were carried out in spontaneously breathing anaesthetized rats (sodium pentobarbitone 60 mg/kg i.p., suplemented with 20 mg/kg i.p.). The cardiorespiratory response evoked by electrical stimulation of the dlPAG (1 ms pulses, 20–50 μA, given at 100 Hz, during 5 s) was analysed before and after the microinjection, within the A5 region, of either kynurenic acid (non-specific glutamate receptor antagonist; 5–10 nmol), DAP-5 (NMDA antagonist; 1 pmol), CNQX (non-NMDA antagonist; 1 pmol) or MCPG (metabotropic antagonist; 0,1 nmol). Kynurenic acid decreased the intensity of both the tachypnoea (p < 0,001) and tachycardia (p < 0,001) induced by dl-PAG stimulation. Blockade of no-NMDA receptors reduced the increase of respiratory frequency, heart rate and pressor response to dl-PAG stimulation (p < 0,01, p < 0,001, p < 0,05 respectively). Blockade of either NMDA or metabotropic receptors reduced the dlPAG-evoked tachycardia and pressor response (p < 0,01; p < 0,05 respectively). These results suggest a neuromodulatory role for A5 region via glutamate neurotransmission of the dlPAG-evoked cardiorespiratory response, confirming the role of the ventrolateral pons in the neuronal circuits involved in respiratory and heart rate control.

BackgroundThere is currently a wide therapeutic arsenal for migraine patients, without a single first-line preventive drug and we choose the different available alternatives taking into account comorbidities, national guidelines, previous treatments and personal experiences.Our objective was to evaluate the differences in the use of migraine treatments between neurologists from different countries.MethodsThis is a multi-centre observational study carried out by neurologists from specialized headache units in seven countries, retrospective with consecutive inclusion of all patients presenting with a migraine diagnosis, over a period of three months.ResultsA total of 734 patients were recruited but only 600 were considered in the analysis in order to homogenize the patient cohorts from countries: 200 Spain (ES), 100 Italy (IT), 85 Russia (RUS), 80 Germany (DE), 60 Portugal (PT), 45 Poland (PL) and 30 Australia (AU). 85.4 % of patients were women with a mean age of 42.6 ± 11.8 years. Considering previous and current preventive treatment, the order of use was: antidepressants (69.3 %), antiepileptic drugs (54.7 %), beta-blockers and antihypertensive drugs (49.7 %), OnabotulinumtoxinA (44.0 %) and others (36.2 %).Statistically significant differences were found between all pharmacological classes: antidepressants were commonly used in all countries, with the exception of Poland (AU: 76.7 %, IT: 71.0 %, DE: 60.0 %, PL: 31.1 %, PT: 71.7 %, RUS: 70.6 %, ES: 78.5 %; p < 0.0001); antiepileptic drugs were more frequently prescribed in Portugal, Australia and Spain (AU: 73.3 %, IT: 40.0 %, DE: 37.5 %, PL: 48.9 %, PT: 85.0 %, RUS: 29.4 % and ES: 69.0 %; p < 0.0001); beta-blockers and antihypertensive drugs were frequently used in all countries except Italy (AU: 60.0 %, IT: 14.0 %, DE: 53.8 %, PL: 48.9 %, PT: 68.3 %, RUS: 49.4 % and ES: 59.0 %; p < 0.0001); BTX-A were predominately used in Spain, Italy and Australia (AU:56.7 %, IT:58.0 %, DE:20.0 %, PL: 42.2 %, PT: 26.7 %, RUS: 24.7 % and ES: 58.5 %; p < 0.0001) and others were most frequently used in Poland (AU: 0.0 %, IT: 19.0 %, DE: 42.5 %, PL: 95.6 %, PT: 31.7 %, RUS: 3.5 % and ES: 49.5 %; p < 0.0001). If only patients without comorbidities are considered (200/600), statistically differences between countries persist in all preventive treatments.ConclusionsThere is heterogeneity in the choice of preventive treatment between different countries. Prospective comparative studies of the different oral and subcutaneous alternatives would help to create a global therapeutic algorithm that would guarantee the best option for our patients.