Planaria: an animal model that integrates development, regeneration and pharmacology

Although planarians are established model organisms in developmental biology and regeneration studies, in the last forty years or so, they have caught the attention of pharmacologists, especially to study the pharmacology of drugs of abuse. This review covers the following topics: some fundamentals of the history of animal models and planarians in biomedical research; an abbreviated story of systematic pharmacology research using planarians as a model organism; an example of how planarians are contributing to the search for compounds against acute cocaine toxicity; an analysis of the number of papers on planarians and pharmacological topics from 1900- 2016; some perspectives on pharmacology in developmental and regeneration studies, arguing in favor of the planarian model as a leading subject for this interdisciplinary area of research, and finally some concluding thoughts

Exploiting the synergism among physical and chemical processes for improving food safety

This article provides an overview of recent published information on the subject of synergistic lethal effects that emerge from the combination of physical and/or chemical processes applied to enhance food safety. Despite important recent advances in non-thermal technologies, the greatest synergistic lethal effects emerge from combining them with traditional, relatively mild heat treatments. The combined application of antimicrobials has shown that their main constituents interact effectively, and great synergistic effects have been described with the capacity of either inhibiting or inactivating pathogens. Moreover, natural antimicrobials are more effective when pathogens are previously damaged sublethally by the application of physical technologies. Such combinations allow for a considerable reduction of treatment intensity and costs, along with a noticeable improvement in food quality and safety

Incubation with a complex orange essential oil leads to evolved mutants with increased resistance and tolerance

Emergence of strains with increased resistance/tolerance to natural antimicrobials was evidenced after cyclic exposure to carvacrol, citral, and (+)-limonene oxide. However, no previous studies have reported the development of resistance and tolerance to complex essential oils (EOs). This study seeks to evaluate the occurrence of Staphylococcus aureus strains resistant and tolerant to a complex orange essential oil (OEO) after prolonged cyclic treatments at low concentrations. Phenotypic characterization of evolved strains revealed an increase of minimum inhibitory and bactericidal concentration for OEO, a better growth fitness in presence of OEO, and an enhanced survival to lethal treatments, compared to wild-type strain. However, no significant differences (p > 0.05) in cross-resistance to antibiotics were observed. Mutations in hepT and accA in evolved strains highlight the important role of oxidative stress in the cell response to OEO, as well as the relevance of the cell membrane in the cell response to these natural antimicrobials. This study demonstrates the emergence of S. aureus strains that are resistant and tolerant to EO (Citrus sinensis). This phenomenon should be taken into account to assure the efficacy of natural antimicrobials in the design of food preservation strategies, in cleaning and disinfection protocols, and in clinical applications against resistant bacteria

Antimicrobial activity of suspensions and nanoemulsions of citral in combination with heat or pulsed electric fields

The application of essential oils in form of nanoemulsions has been proposed as a method to improve their solubility in aqueous solutions, and hence their antimicrobial activity. The objective of this study was to evaluate the antimicrobial activity of citral, applied directly or in combined treatments with heat or pulsed electric fields (PEF), as a function of the inoculation procedure assayed: (i) a simple, vigorous shaking method by vortex agitation (suspension of citral; s-citral) or (ii) the previous preparation of nanoemulsions by the emulsion phase inversion (EPI) method (nanoemulsion of citral; n-citral). n-Citral was more effective in either inhibiting or inactivating Escherichia coli O157:H7 Sakai than s-citral. However, when combined with heat, a greater synergistic effect was observed with s-citral rather than with n-citral, either in lab media (pH 7·0 and 4·0) or apple juice. For instance, while almost 5 log10 cell cycles were inactivated in apple juice after 15 min at 53°C in the presence of 0·1 µl ml-1 of s-citral, the use of n-citral required 30 min. The use of nanoemulsions did not modify the slight synergism observed when citral and mild PEF were combined (150 µs, 30 kV cm-1). Significance and Impact of the Study: The exploration of different delivery systems of antimicrobial compounds such as citral in aqueous food products aids in the establishment of successful combined treatments for food preservation. While at room temperature, citral in form of a nanoemulsion shows a higher antimicrobial activity; its combination with heat would imply a partial loss of the outstanding synergistic lethal effect achieved when added in suspension form. Therefore, the most suitable procedure to magnify the synergism between heat and citral when processing juices would merely require an intense homogenization step prior to the combined treatment

Modified cyclodextrin type and dehydration methods exert a significant effect on the antimicrobial activity of encapsulated carvacrol and thymol

BACKGROUND The antimicrobial activity of essential oils and their constituents has led to increasing interest in using them as natural preservative agents. However, their high sensitivity to light and oxygen, their volatility and their low aqueous solubility are all obstacles to their application in the food, cosmetic or pharmaceutical industries. Encapsulation in cyclodextrins (CDs) is a solution for the application of such essential oils. RESULTS The complexation of carvacrol and thymol with hydroxypropyl (HP)‐α‐, HP‐β‐ and HP‐γ‐CD, the behavior of the solid complexes prepared by freeze‐drying and spray‐drying methods and the antibacterial activity of solid complexes were studied. Kc values of HP‐α‐ and HP‐γ‐CD complexes with carvacrol (118.4 and 365.7 L mol−1) and thymol (112.5 and 239.7 L mol −1) were far lower than those observed for HP‐β‐CD complexes with carvacrol (2268.2 L mol −1) and thymol (881.6 L mol −1). The lower stability of HP‐α‐ and HP‐γ‐CD complexes increased the release of compounds, thereby affecting the antimicrobial activity of carvacrol and thymol to a lesser extent than complexation with HP‐β‐CD, normally used in the encapsulation of carvacrol and thymol. HP‐β‐CD encapsulation of carvacrol and thymol markedly reduced their antimicrobial activity. The freeze‐drying method barely affected the antimicrobial activity of carvacrol and thymol after encapsulation, while spray drying could be considered for the production of solid complexes in combination with the appropriate CD. CONCLUSIONS It was thus demonstrated that HP‐α‐ and HP‐γ‐CD are very suitable alternatives for the encapsulation of carvacrol and thymol with the purpose of preserving their bacteriostatic and bactericidal activities

Nanoemulsions of Mentha piperita L. essential oil in combination with mild heat, pulsed electric fields (PEF) and high hydrostatic pressure (HHP) as an alternative to inactivate Escherichia coli O157: H7 in fruit juices

This study was performed in order to obtain and characterize nanoemulsions of Mentha piperita L. essential oil (n-MPEO) to assess its efficacy in combination with mild heat (MHT) (50, 52, 54¿°C; 10¿min), pulsed electric fields (PEF) (20, 25, 30¿kV/cm; 150¿µs), and high hydrostatic pressure (HHP) (150, 200, 300¿MPa; 15¿min) treatments in causing a 5- log10 reduction in survival counts (from 7 to <2 log10 CFU/mL) of Escherichia coli O157:H7 in guava and mango juices. The droplets of n-MPEO were¿<¿200¿nm and showed good stability for 4¿months at 4¿°C. The n-MPEO at 5.0¿µL/mL displayed a more efficacious long-term antimicrobial activity than suspensions of MPEO (s-MPEO). Combined treatments of s-MPEO or n-MPEO and MHT, PEF, or HHP acted synergistically against E. coli. Nevertheless, combined treatments with n-MPEO showed the same or even higher efficacy than those with s-MPEO (up to 1- additional log10 reduction). Thus, by using nanoemulsions, lower doses of antimicrobial compounds (up to 4 times) or milder MHT (up to 4¿°C), PEF (up to 5¿kV/cm) and HHP (up to 100¿MPa) treatments can be applied, while still guaranteeing the microbial safety of tropical fruit juices

Detection of thermal sublethal injury in escherichia coli via the selective medium plating technique: Mechanisms and improvements

In food preservation, the synergistic combination of different technologies aims to maximize the total lethality of the process and minimize the intensity of each hurdle. This is especially the case when at least one of the treatments can cause sublethal (reparable) injury in a great proportion of the population, so that sublethally injured cells can end up being entirely inactivated by the other hurdle(s). The selective medium plating technique (SMPT) is extensively used to enumerate bacterial sublethal injury after inimical treatments, being sodium chloride added to the recovery medium to detect damaged bacterial envelopes. However, little work has been done to explain the reasons for the inability of sublethally injured cells to outgrow in selective agar media, whereas they are able to grow in non-selective agar. In the present paper, the performance of SMPT on Escherichia coli cells after heat treatments is explored by applying different selective agents in the recovery media, using mutants lacking factors involved in osmoregulation, and also by examining the integrity of the cytoplasmic membrane. In view of the results, the possibility of a specific toxic effect of Na+ as the main mechanism under SMPT was discarded, since the same level of sublethal injury was detected using KCl instead of NaCl. The synthesis of the osmoprotectant trehalose determined the maximum osmotolerance of intact cells to the selective agents, but was not crucial in the quantification of sublethal injury. Moreover, for the first time, the extent of sublethal injury detected via SMPT was directly correlated with the physical loss of integrity of the cell membrane in 99.999% of the initial population. This was achieved through statistical analysis of flow cytometry data using propidium iodide-exclusion technique when that dye was added before thermal treatments. The present work confirms the adequacy of SMPT as a tool for detecting the occurrence and quantity of sublethally injured cells after thermal treatments and thus, for efficiently designing the combination of heat with other preservation techniques. We also propose the study of statistical analysis from flow cytometry data for a more rapid quantification of bacterial sublethal injury in a broad detection range

PAMP-triggered immunity against Pseudomonas syringae involves microRNA-mediated regulation of several uncharacterized R genes

Two main types of noncoding small RNA molecules have been found in plants: microRNAs (miRNAs) and small interfering RNAs (siRNAs). They differ in their biogenesis and mode of action, but share similar sizes (20-24 nt). Their precursors are processed by Dicer-Like RNase III (dcl) proteins present in Arabidopsis thaliana, and in their mature form can act as negative regulators of gene expression, being involved in a vast array of plant processes, including plant development, genomic integrity or response to stress. Small-RNA mediated regulation can occurs at transcriptional level (TGS) or at post-transcriptional level (PTGS). In recent years, the role of gene silencing in the regulation of expression of genes related to plant defence responses against bacterial pathogens is becoming clearer. Comparisons carried out in our lab between the expression profiles of different mutants affected in gene silencing, and plants challenged with Pseudomonas syringae pathovar tomato DC3000, led us to identify a set of uncharacterized R genes, belonging to the TIR-NBS-LRR gene family, differentially expressed in these conditions. Through the use of bioinformatics tools, we found a miRNA* of 22 nt putatively responsible for down-regulating expression of these R genes through the generation of siRNAs. We have also found that the corresponding pri-miRNA is down-regulated after PAMP-perception in a SA-dependent manner. We also demonstrate that plants with altered levels of miRNA* (knockdown lines or overexpression lines) exhibit altered PTI-associated phenotypes, suggesting a role for this miRNA* in this defence response against bacteria. In addition we identify one of the target genes as a negative regulator of defence response against Pseudomonas syringae.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. MINECO, FEDE