FOXO1 regulates wound-healing responses in human gingival fibroblasts.

BACKGROUND AND OBJECTIVE: Forkhead box-O 1 (FOXO1) is a transcription factor actively involved in oral wound healing at the epithelial barrier. However, less is known regarding the role of FOXO1 during the tissue repair response in the connective tissue compartment. This study explored the involvement of FOXO1 in the modulation of fibroblast activity related to wound healing. METHODS: Primary cultures of human gingival fibroblasts were obtained from four healthy young donors. Myofibroblastic differentiation, collagen gel contraction, cell migration, cell spreading, and integrin activation were evaluated in the presence or absence of a FOXO1 inhibitor (AS1842856). Variations in mRNA and proteins of interest were evaluated through qRT-PCR and western blot, respectively. Distribution of actin, α-smooth muscle actin, and ß1 integrin was evaluated using immunofluorescence. FOXO1 and TGF-ß1 expression in gingival wound healing was assessed by immunohistochemistry in gingival wounds performed in C57BL/6 mice. Images were analyzed using ImageJ/Fiji. ANOVA or Kruskal-Wallis test followed by Tukey's or Dunn's post-hoc test was performed. All data are expressed as mean ± SD. p < .05 was considered statistically significant. RESULTS: FOXO1 inhibition caused a decrease in the expression of the myofibroblastic marker α-SMA along with a reduction in fibronectin, type I collagen, TGF-ß1, and ß1 integrin mRNA level. The FOXO1 inhibitor also caused decreases in cell migration, cell spreading, collagen gel contraction, and ß1 integrin activation. FOXO1 and TGF-ß1 were prominently expressed in gingival wounds in fibroblastic cells located at the wound bed. CONCLUSION: The present study indicates that FOXO1 plays an important role in the modulation of several wound-healing functions in gingival fibroblast. Moreover, our findings reveal an important regulatory role for FOXO1 on the differentiation of gingival myofibroblasts, the regulation of cell migration, and collagen contraction, all these functions being critical during tissue repair and fibrosis.

Atypical COVID-19 presentation with Budd-Chiari syndrome leading to an outbreak in the emergency department.

We described a case report of a 50 years-old-woman admitted to the emergency department with abdominal pain associated with febrile hepatosplenomegaly with the final diagnosis of suprahepatic vein thrombosis secondary to COVID-19. Initially, this patient stayed out of a private room because of this atypical presentation and caused a COVID-19 outbreak in the emergency department.

Insecticidal, Repellent and Antifeedant Activity of Essential Oils from Blepharocalyx cruckshanksii (Hook. & Arn.) Nied. Leaves and Pilgerodendron uviferum (D. Don) Florin Heartwood against Horn Flies, Haematobia irritans (Diptera: Muscidae).

Haematobia irritans is a cosmopolitan obligate blood-feeding ectoparasite of cattle and is the major global pest of livestock production. Currently, H. irritans management is largely dependent on broad-spectrum pesticides, which has led to the development of insecticide resistance. Thus, alternative control methods are needed. Essential oils have been studied as an alternative due to their wide spectrum of biological activities against insects. Thus, the main aim of this study was to evaluate the insecticidal, repellent and antifeedant activity of the essential oils from Blepharocalyx cruckshanksii leaves and Pilgerodendron uviferum heartwood against horn flies in laboratory conditions. The composition of the essential oils was analyzed using gas chromatography coupled to mass spectrometry. Accordingly, α-pinene (36.50%) and limonene (20.50%) were the principal components of the B. cruckchanksii essential oil, and δ-cadinol (24.16%), cubenol (22.64%), 15-copaenol (15.46%) and δ-cadinene (10.81%) were the most abundant compounds in the P. uviferum essential oil. Mortality of flies and feeding behavior were evaluated by non-choice tests, and olfactory response was evaluated using a Y-tube olfactometer. Both essential oils were toxic to horn flies, with LC50 values for B. cruckchanksii essential oil of 3.58 µL L-1 air at 4 h, and for P. uviferum essential oil of 9.41 µL L-1 air and 1.02 µL L-1 air at 1 and 4 h, respectively. Moreover, the essential oils exhibited spatial repellency in the olfactometer using only 10 µg of each oil, and these significantly reduced the horn fly feeding at all doses evaluated. Although further laboratory and field studies related to the insectistatic and insecticide properties of these essential oils against H. irritans are necessary, B. cruckshanksii leaves and P. uviferum heartwood essential oils are promising candidates for horn fly management.

Antifeedant Effects and Repellent Activity of Loline Alkaloids from Endophyte-Infected Tall Fescue against Horn Flies, Haematobia irritans (Diptera: Muscidae).

Haematobia irritans is an obligate bloodsucking ectoparasite of cattle and is the global major pest of livestock production. Currently, H. irritans management is largely dependent upon broad-spectrum pesticides, which lately has led to the development of insecticide resistance. Thus, alternative control methods are necessary. Endophyte-infected grasses have been studied as an alternative due to their capability to biosynthesize alkaloids associated with anti-insect activities. Thus, the main aim of this study was to evaluate the antifeedant and repellent activity of lolines obtained from endophyte-infected tall fescue against H. irritans adults in laboratory conditions. The alkaloid extract (ALKE) was obtained by acid-base extraction. N-formyl loline (NFL) and N-acetyl loline (NAL) were isolated by preparative thin layer chromatography (pTLC) and column chromatography (CC), and the loline was prepared by acid hydrolysis of a NFL/NAL mixture. Loline identification was performed by gas chromatography coupled to mass spectrometry (GC/MS). Feeding behavior was evaluated by a non-choice test, and olfactory response was evaluated using a Y-tube olfactometer. Accordingly, all samples showed antifeedant activities. NFL was the most antifeedant compound at 0.5 µg/µL and 1.0 µg/µL, and it was statistically equal to NAL but different to loline; however, NAL was not statistically different to loline. NFL and NAL at 0.25 µg/µL were more active than loline. All samples except loline exhibited spatial repellency in the olfactometer. Thus, the little or non-adverse effects for cattle and beneficial activities of those lolines make them suitable candidates for horn fly management.

Uncoupled inflammatory, proliferative, and cytoskeletal responses in senescent human gingival fibroblasts.

BACKGROUND AND OBJECTIVE: Aging is characterized by a decline in tissue structure and function that may be explained by the development of cellular senescence. However, the acquisition of specific phenotypic responses in senescent gingival fibroblasts is still poorly understood. Here, we have analyzed whether proliferation of primary cultures of human gingival fibroblasts may affect different cell functions relevant to cellular senescence and tissue deterioration. METHODS: Human gingival fibroblasts from five young donors were expanded until cellular senescence was achieved. Cellular senescence was evaluated by determining modifications in cell size, cell proliferation, p16 and p21 mRNA levels, H2Ax phosphorylation, cell viability, and senescence-associated beta-galactosidase staining. Inflammation was evaluated by analyzing the secretion of cytokines and nuclear translocation of NF-κB. Collagen remodeling was evaluated using a collagen gel contraction assay. Immunofluorescence and confocal microscopy were used to determine changes in the localization of the cytoskeletal proteins. Data analysis was performed to identify changes between cultures of the same donor at early and late passages using the paired sample t test or the Wilcoxon matched-pairs signed-rank test. RESULTS: Late passage cells showed changes compatible with cellular senescence that included increased cell size, reduced cell proliferation, staining for SA-beta gal, phosphorylated H2Ax, and increased p16 and p21 mRNA levels. Late passage cells showed a decrease in collagen contraction and reduced co-localization between the cytoskeletal proteins actin and vinculin. Importantly, late passage cells neither demonstrated changes in the secretion of inflammatory cytokines nor NF-κB activation. CONCLUSION: Our results imply that cytoskeletal changes and inhibition of cell proliferation represent early modifications in the structure and function of senescent gingival fibroblasts that are not coupled with the acquisition of an inflammatory phenotype. Further studies are needed to clarify the impact of different senescence stages during aging of the periodontium.

Antifeedant Effects of Essential Oil, Extracts, and Isolated Sesquiterpenes from Pilgerodendron uviferum (D. Don) Florin Heartwood on Red Clover Borer Hylastinus obscurus (Coleoptera: Curculionidae).

The beetle Hylastinus obscurus Marsham (Coleoptera: Curculionidae), endemic to Europe and Northern Africa, is one of the most important red clover pests in Chile. As commercial insecticides are less effective against this pest, plant secondary metabolites have been considered as an alternative for its control. Here, we have investigated the chemical composition of essential oil (EO), petroleum ether extract (PEE), and dichloromethane extract (DCME) from Pilgerodendron uviferum heartwood. Additionally, the effects of EO and extracts on the feeding behavior (% of weight shift) of H. obscurus have been evaluated. The composition of EO, PEE, and DCME were analyzed using gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The results showed the presence of a similar mixture of sesquiterpenes in the essential oil and in both of the extracts, which accounted for circa 60% of the total mixture. Sesquiterpenes were further isolated using chromatographic methods and were structurally characterized by optical rotation, GC⁻MS, FTIR, and 1D and 2D NMR experiments. The physicochemical properties of the isolated sesquiterpenes, including lipophilicity and vapor pressure, were also determined. The sesquiterpenes were identified as the following: (-)-trans-calamenene (1), cadalene (2), (-)-cubenol (3), (-)-epi-cubenol (4), (-)-torreyol (5), and (-)-15-copaenol (6). The antifeedant activity of EO, extracts, and isolated sesquiterpenes were evaluated using artificial diets in a non-choice test. Relative to the control, the EO, DCME extract, and the isolated sesquiterpenes, namely, (-)-trans-calamenene (1), cadalene (2), and (5) torreyol, were found to be the most effective treatments against H. obscurus. Our study showed that the compounds occurring in P. uviferum heartwood were effective in reducing the adult growth of H. obscurus. The physicochemical properties of the isolated sesquiterpenes might have been associated with antifeedant effects.

Repellent Activity of the Essential Oil from the Heartwood of Pilgerodendron uviferum (D. Don) Florin against Aegorhinus superciliosus (Coleoptera: Curculionidae).

The weevil Aegorhinus superciliosus Guérin (Coleoptera: Curculionidae), which is endemic to Central-Southern Chile and Argentina, is one of the major berry pests in Chile and the most important pest in the La Araucanía Region (38°44'9″S, 72°35'25″W). Due to the poor effectiveness and problems surrounding the implementation of the traditional control methods using organophosphate and carbamate insecticides, new strategies for controlling this pest are needed. In this communication, we evaluated the behavioral responses of male and female A. superciliosus to volatile compounds released from the essential oil (EO) obtained from the heartwood of Pilgerodendron uviferum (D. Don) Florin using olfactometric bioassays. The composition of the EO was analyzed using gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). According to these analyses, δ-cadinol (24.16%), cubenol (22.64%), 15-copaenol (15.46%) and δ-cadinene (10.81%) were the principal components of the EO. The Pilgerodendron uviferum EO, which is almost exclusively composed of sesquiterpenes (99.5%), exhibited a repellent effect against A. superciliosus adults, regardless of the sex or concentration used (56.6 mg/cm³ and 1.58 × 10(-2) mg/cm³). The EO has low volatility and greater persistence than the EOs composed of monoterpenes and is considered a good model in the search for raspberry weevil repellents.

Formulation of essential oils-loaded solid lipid nanoparticles-based chitosan/PVA hydrogels to control the growth of Botrytis cinerea and Penicillium expansum.

Botrytis cinerea and Penicillium expansum are phytopathogenic fungi that produce the deterioration of fruits. Thus, essential oil (EO) has emerged as a sustainable strategy to minimize the use of synthetic fungicides, but their volatility and scarce solubility restrict their application. This study proposes the EO of Oreganum vulgare and Thymus vulgaris-loaded solid lipid nanoparticles (SLN) based chitosan/PVA hydrogels to reduce the infestation of fungi phytopathogen. EO of O. vulgare and T. vulgaris-loaded SLN had a good homogeneity (0.21-0.35) and stability (-28.8 to -33.0 mV) with a mean size of 180.4-188.4 nm. The optimization of EO-loaded SLN showed that the encapsulation of 800 and 1200 µL L-1 of EO of O vulgare and T. vulgaris had the best particle size. EO-loaded SLN significantly reduced the mycelial growth and spore germination of both fungi pathogen. EO-loaded SLN into hydrogels showed appropriate physicochemical characteristics to apply under environmental conditions. Furthermore, rheological analyses evidenced that hydrogels had solid-like characteristics and elastic behavior. EO-loaded SLN-based hydrogels inhibited the spore germination in B. cinerea (80.9 %) and P. expansum (55.7 %). These results show that SLN and hydrogels are eco-friendly strategies for applying EO with antifungal activity.

Evolution of the vertebrate bone matrix: an expression analysis of the network forming collagen paralogues in amphibian osteoblasts.

The emergence of vertebrates is closely associated to the evolution of mineralized bone tissue. However, the molecular basis underlying the origin and subsequent diversification of the skeletal mineralized matrix is still poorly understood. One efficient way to tackle this issue is to compare the expression, between vertebrate species, of osteoblastic genes coding for bone matrix proteins. In this work, we have focused on the evolution of the network forming collagen family which contains the Col8a1, Col8a2, and Col10a1 genes. Both phylogeny and synteny reveal that these three paralogues are vertebrate-specific and derive from two independent duplications in the vertebrate lineage. To shed light on the evolution of this family, we have analyzed the osteoblastic expression of the network forming collagens in endochondral and intramembraneous skeletal elements of the amphibian Xenopus tropicalis. Remarkably, we find that amphibian osteoblasts express Col10a1, a gene strongly expressed in osteoblasts in actinopterygians but not in amniotes. In addition, while Col8a1 is known to be robustly expressed in mammalian osteoblasts, the expression levels of its amphibian orthologue are dramatically reduced. Our work reveals that while a skeletal expression of network forming collagen members is widespread throughout vertebrates, osteoblasts from divergent vertebrate lineages express different combinations of network forming collagen paralogues.

Effect of lithium on the properties of a liquid crystal formed by sodium dodecylsulphate and decanol in aqueous solution.

Understanding the molecular interactions that rule the physicochemical properties of molecular assemblies is of particular interest when trying to explain the behavior of much more complicated systems, such as the cell membranes. This work was devoted to study a discotic nematic lyotropic liquid crystal, formed by sodium dodecylsulphate (3% SDS-d25) and decanol (20% DeOH-α-d2), dissolved in aqueous solutions (0.1% D2O) of Na2SO4 or Li2SO4. The average size of the aggregates was estimated using fluorescence quenching experiments, and their dynamics were studied by measuring the (2)H-NMR quadrupole splitting (ΔνQ) and the longitudinal relaxation times (T1) of the deuterated species. To provide an atomic insight into these assemblies, molecular dynamics simulations of the systems were carried out with atomic detail. As a previous step in this study, a reparameterization of the standard GROMOS 87 force field was required to perform the equilibrated simulations and to prevent instabilities emerging during the simulations. Finally, an excellent agreement between simulation and experimental data was obtained. In addition, variations in the long range electrostatic interactions at the aggregate/solution interface, the orientation and the reorientational relaxation time of the water dipole, the translational diffusion coefficient of sodium ions, and the amphiphile-counterion coordination associated with the presence of Li(+) in the solution were other key aspects investigated to explain the variation in the quadrupole splittings (ΔνQ) in the presence of lithium in solution.

Herbivory Damage Increased VOCs in Wild Relatives of Murtilla Plants Compared to Their First Offspring.

Murtilla (Ugni molinae) is a shrub native to Chile that has undergone an incipient domestication process aimed at increasing its productivity. The reduction in intrinsic chemical defenses due to the domestication process has resulted in a decrease in the plant's ability to defend itself against mechanical or insect damage. In response to this damage, plants release volatile organic compounds (VOCs) as a means of defense. To understand how domestication has impacted the production of VOCs in the first offspring of murtilla, we hypothesized that their levels would be reduced due to the induction of mechanical and herbivore damage. To test this hypothesis, we collected VOCs from four offspring ecotypes and three wild relatives of murtilla. We induced mechanical and herbivore damage in the plants and then enclosed them in a glass chamber, where we captured the VOCs. We identified 12 compounds using GC-MS. Our results showed that wild relative ecotypes had a higher VOC release rate of 624.6 µg/cm2/day. Herbivore damage was the treatment that produced the highest release of VOCs, with 439.3 µg/cm2/day in wild relatives. These findings suggest that herbivory triggers defenses through the emission of VOCs, and that domestication has influenced the production of these compounds in murtilla. Overall, this study contributes to bridging the gap in the incipient domestication history of murtilla and highlights the importance of considering the impact of domestication on a plant's chemical defenses.

The efficient activity of plant essential oils for inhibiting Botrytis cinerea and Penicillium expansum: Mechanistic insights into antifungal activity.

Botrytis cinerea and Penicillium expansum produce deterioration in fruit quality, causing losses to the food industry. Thus, plant essential oils (EOs) have been proposed as a sustainable alternative for minimizing the application of synthetic fungicides due to their broad-spectrum antifungal properties. This study investigated the efficacy of five EOs in suppressing the growth of B. cinerea and P. expansum and their potential antifungal mechanisms. EOs of Mentha × piperita L., Origanum vulgare L., Thymus vulgaris L., Eucalyptus globules Labill., and Lavandula angustifolia Mill., were screened for both fungi. The results showed that the EO of T. vulgaris and O. vulgare were the most efficient in inhibiting the growth of B. cinerea and P. expansum. The concentration increase of all EO tested increased fungi growth inhibition. Exposure of fungi to EOs of T. vulgaris and O. vulgare increased the pH and the release of constituents absorbing 260 nm and soluble proteins, reflecting membrane permeability alterations. Fluorescence microscopic examination revealed that tested EOs produce structural alteration in cell wall component deposition, decreasing the hypha width. Moreover, propidium iodide and Calcein-AM stains evidenced the loss of membrane integrity and reduced cell viability of fungi treated with EOs. Fungi treated with EOs decreased the mitochondria activity and the respiratory process. Therefore, these EOs are effective antifungal agents against B. cinerea and P. expansum, which is attributed to changes in the cell wall structure, the breakdown of the cell membrane, and the alteration of the mitochondrial activity.

The Impact of 2-Ketones Released from Solid Lipid Nanoparticles on Growth Modulation and Antioxidant System of Lactuca sativa.

2-Ketones are signal molecules reported as plant growth stimulators, but their applications in vegetables have yet to be achieved. Solid lipid nanoparticles (SLNs) emerge as a relevant nanocarrier to develop formulations for the controlled release of 2-ketones. In this sense, seedlings of Lactuca sativa exposed to 125, 375, and 500 µL L-1 of encapsulated 2-nonanone and 2-tridecanone into SLNs were evaluated under controlled conditions. SLNs evidenced a spherical shape with a size of 230 nm. A controlled release of encapsulated doses of 2-nonanone and 2-tridecanone was observed, where a greater release was observed as the encapsulated dose of the compound increased. Root development was strongly stimulated mainly by 2-tridecanone and leaf area (25-32%) by 2-nonanone. Chlorophyll content increased by 15.8% with exposure to 500 µL L-1 of 2-nonanone, and carotenoid concentration was maintained with 2-nonanone. Antioxidant capacity decreased (13-62.7%) in L. sativa treated with 2-ketones, but the total phenol concentration strongly increased in seedlings exposed to some doses of 2-ketones. 2-Tridecanone strongly modulates the enzymatic activities associated with the scavenging of H2O2 at intra- and extracellular levels. In conclusion, 2-ketones released from SLNs modulated the growth and the antioxidant system of L. sativa, depending on the dose released.

The Essential Oil of Petroselinum crispum (Mill) Fuss Seeds from Peru: Phytotoxic Activity and In Silico Evaluation on the Target Enzyme of the Glyphosate Herbicide.

Petroselinum crispum (Mill) Fuss is an aromatic plant belonging to the Apiaceae family and used in gastronomy as a spice. Several studies have been developed in leaves but studies are limited in seeds, especially the essential oils obtained from seeds. The aim of this study was to determine the phytochemical profile of the volatile compounds of this essential oil by gas-chromatography-mass spectrometry (GC-MS) in order to evaluate its phytotoxic activity on Lactuca sativa seeds and to carry out an in silico analysis on the target enzyme of the herbicide glyphosate 5-enolpyruvylshikimate 3-phosphate synthase (EPSP). The essential oil was obtained by steam distillation for two hours and then was injected into a GC-MS, the phytotoxic assay was carried out on Lactuca seeds and the in silico evaluation on the EPSP synthase focused on the volatile compounds similar to glyphosate, docking analysis, and molecular dynamics to establish the protein-ligand stability of the most active molecule. The chromatographic analysis revealed 47 compounds, predominated by three compounds with the most abundant percentage in the total content (1,3,8-ρ-menthatriene (22.59%); apiole (22.41%); and ß-phellandrene (15.02%)). The phytotoxic activity demonstrated that the essential oil had a high activity at 5% against L. sativa seed germination, inhibition of root length, and hypocotyl length, which is comparable to 2% glyphosate. The molecular docking on EPSP synthase revealed that trans-p-menth-6-en-2,8-diol had a high affinity with the enzyme EPSP synthase and a better stability during the molecular dynamic. According to the results, the essential oil of P. crispum seeds presented a phytotoxic activity and might be useful as a bioherbicide agent against weeds.

A temporary immersion system for mass micropropagation of pitahaya (Hylocereus undatus).

Scaling-up techniques in temporary immersion systems are an alternative for commercial micropropagation. In vitro propagation of pitahaya (Hylocereus undatus) using temporary immersion with liquid culture medium improves micropropagation efficiency compared to the conventional method in semisolid medium. The objective of this work was to evaluate the efficiency of traditional culture and temporary immersion during micropropagation of pitahaya to facilitate the rapid establishment of new commercial plantations of high genetic and phytosanitary quality. Semisolid culture, liquid media in partial immersion and temporary immersion in an Ebb-and-Flow bioreactor were evaluated. Also, in temporary immersion, different immersion frequencies (every 4, 8, 12, and 16 h) and culture densities (5, 10, 15 and 20 explants per bioreactor) were evaluated. For the multiplication stage, new shoots and length were recorded per explant at 45 d of in vitro culture and in the acclimatization stage, the survival percentage was determined at 30 d of greenhouse cultivation. A temporary immersion of 2 min every 4 h and 15 explants per bioreactor was the best culture system, obtaining on average 10.7 shoots per explant with a length of 1.9 cm. No significant differences were observed among treatments during acclimatization, obtaining survival percentages of 98%-100%. This study reports for the first time a protocol for scaling-up techniques in temporary immersion for commercial micropropagation of pitahaya (and for any species of the Cactaceae family) and its establishment in a productive plantation.

Two families of Xenopus tropicalis skeletal genes display well-conserved expression patterns with mammals in spite of their highly divergent regulatory regions.

The origin of bone and cartilage, and their subsequent diversification in specific vertebrate lineages, is intimately linked to the precise transcriptional control of genes involved in matrix mineralization. It is not yet clear, however, to which extent the osteoblasts, osteocytes, and chondrocytes of each of the major vertebrate groups express similar sets of genes. In this study we have focused on the evolution of two independent families of genes that code for extracellular matrix components of the skeleton and that include secreted protein, acidic, cysteine-rich (SPARC), bone sialoprotein (BSP) and dentin matrix protein 1 (DMP1) paralogues, and the osteocalcin (OC) and matrix gla protein (MGP) paralogues. Analyzing developing Xenopus tropicalis skeletal elements, we show that the expression patterns of these genes are well conserved with mammals. The fact that only a few osteoblasts express DMP1, while only some osteocytes express SPARC and BSP, reveals a significant degree of molecular heterogeneity for these two populations of X. tropicalis cells, similarly to what has been described in mouse. Although the cis-regulatory modules (CRM) of the mammalian OC, DMP1, and BSP orthologs have been functionally characterized, we found no evidence of sequence similarity between these regions and the X. tropicalis genome. Furthermore, these regulatory elements evolve rapidly, as they are only poorly conserved between human and rodents. Therefore, the SPARC/DMP1/BSP and the OC/MGP families provide a good paradigm to study how transcriptional output can be maintained in skeletal cells despite extensive sequence divergence of CRM.

Labdanes: antifungal compounds isolates from the resinous exudate of Madia sativa against Phytophthora cinnamomi Rands.

In this study, resinous exudate from Madia sativa was analyzed by GC-MS. The major bioactive compounds 13,14,15-trihydroxylabd-7-ene (14) and 3,14,15-trihydroxylabd-8-ene (15) were isolated and their structures were determined by NMR. The antifungal activity of the resinous exudate and the labdanes compounds was evaluated using the inhibitory effects on the mycelial growth of plant pathogen Phytophthora cinnamomi causing root rot of various crops. The evaluation of the resinous exudate showed no inhibition over 50% at 75 mg/L, while compound 15 had the stronger effect on the myceliar growth of P. cinnamomi, with a 94.6% inhibition at 175 mg/L. The same way, the mixture of both compounds in equal parts did not show a synergistic effect but showed similar percentages of mycelial growth inhibition from 25 mg/L with respect to the compounds separately.

Evaluation of Drimys winteri (Canelo) Essential Oil as Insecticide against Acanthoscelides obtectus (Coleoptera: Bruchidae) and Aegorhinus superciliosus (Coleoptera: Curculionidae).

Adverse effects caused by synthetic pesticides have increased interest in plant-derived insecticidal compounds, in particular essential oils, as a more compatible and ecofriendly alternative for pest control of economic importance. For this reason, the essential oil isolated from leaves and shoots of Drimys winteri (J.R. Forster & G. Forster)-also named canelo (CEO)-was investigated for its chemical profile and insecticidal action against Acanthoscelides obtectus (Say)-one of the most important post-harvest pests of dry beans in the world-and Aegorhinus superciliosus (Guérin)-a significant pest of fruit trees in Chile. The analysis by gas chromatography, paired with mass spectrometry (GC/MS) determined 56 compounds, corresponding to 92.28% of the detected compounds. Elemol (13.54%), γ-eudesmol (11.42%), ß-eudesmol (8.49%), α-eudesmol (6.39%), α-pinene (7.92%) and ß-pinene (5.17%) were the most abundant. Regarding the bioactivity of the CEO, the results demonstrated toxicological effects against A. obtectus. A concentration of 158.3 µL L-1 had a mortality rate of 94% after 24 h exposure. The LC50 and LC90 values at 24 h were 60.1 and 163.0 µL L-1. Moreover, behavioral bioassays showed a repellent effect against A. superciliosus with a dose of one microliter of CEO. Both sexes of the raspberry weevil stayed for very short times in the treated area with the oil (<0.8 min), showing a homogeneous repellency in the species. The overall data suggest that canelo leaves and shoots essential oil has an insecticide effect and is worth exploring to better understand the synergistic relationship between the compounds present in the essential oil.

Essential Oil, Extracts, and Sesquiterpenes Obtained From the Heartwood of Pilgerodendron uviferum Act as Potential Inhibitors of the Staphylococcus aureus NorA Multidrug Efflux Pump.

Staphylococcus aureus is a serious human pathogen that is highly adaptive to environmental conditions and rapidly develops antibiotic resistance. The use of efflux pumps to reduce antibiotic concentrations at the intracellular level is one of the main mechanisms by which bacteria develop antibiotic resistance. The management of efflux pumps, specifically NorA, which is expressed by S. aureus strains, is a valuable strategy for restoring susceptibility in strains resistant to antibacterial agents. In recent years, many studies have focused on searching for natural substances that can reverse efflux pump-mediated resistance in S. aureus. Extracts and compounds obtained from plants can be efficient efflux pump inhibitors (EPIs) and represent a potentially patient-friendly strategy for controlling S. aureus. In the present study, we evaluated the ability of essential oils, petroleum ether extracts, dichloromethane extract (DCME) and six compounds isolated from the heartwood of Pilgerodendron uviferum (Cupressaceae) and two synthetic derivatives to inhibit efflux in NorA pumps in the following three S. aureus strains: K2378, which overexpressed the norA gene (norA++), K1902 (norA-deleted, ΔnorA) and the parental strain, NCTC 8325-4. Efflux activity was evaluated using a fluorometric method that measured the accumulation of the universal efflux pump substrate ethidium bromide (EtBr). Only DCME and the compounds 15-copaenol and epi-cubenol inhibited EtBr efflux by K2378. Even the lowest concentration of 15-copaenol exhibited a stronger inhibitory effect than carbonyl cyanide m-chlorophenyl hydrazone on EtBr efflux by K2378. 15-copaenal only showed inhibition of EtBr efflux in K2378 cells at 125 µg/mL, but not superior to the control inhibitor and 15-copaenyl acetate exerted no intrinsic EPI activity against K2378. Fractional inhibitory concentration index (FICI) values obtained in the checkerboard assays, indicated that all combinations between DCME, epi-cubenol and 15-copaenol, and tested antibiotics showed a synergistic effect in wild type, norA ++ and ΔnorA strains. Moreover, those were not toxic for the HeLa cell line at concentrations in which the synergistic effect and inhibitory activity of efflux pumps was determined. Other extracts and compounds obtained from P. uviferum did not display EtBr efflux-inhibiting activity against the evaluated S. aureus strains.

KDEL receptor regulates secretion by lysosome relocation- and autophagy-dependent modulation of lipid-droplet turnover.

Inter-organelle signalling has essential roles in cell physiology encompassing cell metabolism, aging and temporal adaptation to external and internal perturbations. How such signalling coordinates different organelle functions within adaptive responses remains unknown. Membrane traffic is a fundamental process in which membrane fluxes need to be sensed for the adjustment of cellular requirements and homeostasis. Studying endoplasmic reticulum-to-Golgi trafficking, we found that Golgi-based, KDEL receptor-dependent signalling promotes lysosome repositioning to the perinuclear area, involving a complex process intertwined to autophagy, lipid-droplet turnover and Golgi-mediated secretion that engages the microtubule motor protein dynein-LRB1 and the autophagy cargo receptor p62/SQSTM1. This process, here named 'traffic-induced degradation response for secretion' (TIDeRS) discloses a cellular mechanism by which nutrient and membrane sensing machineries cooperate to sustain Golgi-dependent protein secretion.