The Protective Effect of Lithium Against Rotenone may be Evolutionarily Conserved: Evidence from Eisenia fetida, a Primitive Animal with a Ganglionic Brain.

Chronic exposure to stress is a non-adaptive situation that is associated with mitochondrial dysfunction and the accumulation of reactive oxygen species (ROS), especially superoxide anion (SA). This accumulation of ROS produces damage-associated molecular patterns (DAMPs), which activate chronic inflammatory states and behavioral changes found in several mood disorders. In a previous study, we observed that an imbalance of SA triggered by rotenone (Ro) exposure caused evolutionarily conserved oxi-inflammatory disturbances and behavioral changes in Eisenia fetida earthworms. These results supported our hypothesis that SA imbalance triggered by Ro exposure could be attenuated by lithium carbonate (LC), which has anti-inflammatory properties. The initial protocol exposed earthworms to Ro (30 nM) and four different LC concentrations. LC at a concentration of 12.85 mg/L decreased SA and nitric oxide (NO) levels and was chosen to perform complementary assays: (1) neuromuscular damage evaluated by optical and scanning electron microscopy (SEM), (2) innate immune inefficiency by analysis of Eisenia spp. extracellular neutrophil traps (eNETs), and (3) behavioral changes. Gene expression was also evaluated involving mitochondrial (COII, ND1), inflammatory (EaTLR, AMP), and neuronal transmission (nAchR α5). LC attenuated the high melanized deposits in the circular musculature, fiber disarrangement, destruction of secretory glands, immune inefficiency, and impulsive behavior pattern triggered by Ro exposure. However, the effects of LC and Ro on gene expression were more heterogeneous. In summary, SA imbalance, potentially associated with mitochondrial dysfunction, appears to be an evolutionary component triggering oxidative, inflammatory, and behavioral changes observed in psychiatric disorders that are inhibited by LC exposure.

Superoxide-imbalance Pharmacologically Induced by Rotenone Triggers Behavioral, Neural, and Inflammatory Alterations in the Eisenia fetida Earthworm.

BACKGROUND: Some studies have suggested that mitochondrial dysfunction and a superoxide imbalance could increase susceptibility to chronic stressful events, contributing to the establishment of chronic inflammation and the development of mood disorders. The mitochondrial superoxide imbalance induced by some molecules, such as rotenone, could be evolutionarily conserved, causing behavioral, immune, and neurological alterations in animals with a primitive central nervous system. OBJECTIVE: Behavioral, immune, and histological markers were analyzed in Eisenia fetida earthworms chronically exposed to rotenone for 14 days. METHODS: Earthworms were placed in artificial soil containing 30 nM of rotenone distributed into a plastic cup that allowed the earthworms to leave and return freely into the ground. Since these organisms prefer to be buried, the model predicted that the earthworms would necessarily have to return to the rotenone-contaminated medium, creating a stressful condition. The effect on survival behavior in the immune and histological body wall and ventral nervous ganglia (VNG) structures, as well as gene expression related to inflammation and mitochondrial and neuromuscular changes. RESULTS: Rotenone-induced loss of earthworm escape behavior and immune alterations indicated a chronic inflammatory state. Some histological changes in the body wall and VNG indicated a possible earthworm reaction aimed at protecting against rotenone. Overexpression of the nicotinic acetylcholine receptor gene (nAChR α5) in neural tissues could also help earthworms reduce the degenerative effects of rotenone on dopaminergic neurons. CONCLUSION: These data suggest that mitochondrial dysfunction could be an evolutionarily conserved element that induces inflammatory and behavioral changes related to chronic stress.

Amazonian Guarana- and Açai-Conjugated Extracts Improve Scratched Fibroblast Healing and Eisenia fetida Surgical Tail Amputation by Modulating Oxidative Metabolism.

Background: Previous studies have suggested that guarana (Paullinia cupana) and açai (Euterpe oleracea) have antioxidant, anti-inflammatory, and proliferative properties, indicating their potential therapeutic action in wound healing. We produced a conjugated guarana-açai (GA) extract and tested its healing action on earthworms (Eisenia fetida) subjected to tail amputation by surgical incision. Methods: Extract from roasted guarana seeds and fresh açai seed berries was produced. The antioxidant and genoprotective capacity of GA extract was tested. The concentration with the most remarkable healing potential was used in subsequent tests. The last three posterior segments of the clitellate earthworm tail reared under standardized conditions were surgically amputated. Next, topical PBS or GA extract was applied to the surgical wound. The rate of cell migration and tissue regeneration at the local wound site was histologically evaluated after the procedure. Expression of the SOX4 gene that acts in epithelial-to-mesenchymal transition was determined by RT-qPCR. Results: Sixteen bioactive molecules, including some previously described substances, were identified. All tested concentrations exhibited antioxidant and genoprotective effects. The GA extract accelerated the healing processes as observed through macroscopic and histological analyses and increased expression of SOX4. Conclusion: The GA extract has a potential role in the healing of surgically induced wounds.

Vitamin C as a shelf-life extender in liposomes

Abstract The objective of this study was to evaluate the influence of vitamin C (VC) on the stability of stored liposomes under different climatic conditions. Liposomal formulations containing 1 mg/mL of VC (LIP-VC) and blank formulations (LIP-B) were prepared by the reverse-phase evaporation method. After preparation, they were characterized according to their refractive index, average vesicle diameter, polydispersity index (PDI), zeta potential, pH, content, encapsulation efficiency (EE%), morphology, stability and antioxidant activity. For stability, LIP-VC and LIP-B were stored in different climatic conditions (4 °C, 25 °C and 40 °C) for 30 days. The LIP-VC presented 1.3365 refractive index, 161 nm of mean diameter, 0.231 PDI, -7.3 mV zeta potential, 3.2 pH, 19.4% EE%, spherical morphology, 1 mg/mL of VC content, and antioxidant activity of 12 and 11.4 μmol of TE/mL for the radical DPPH and ABTS+, respectively. During stability, the LIP-B stored in 40 °C showed an instability in the parameters: PDI, vesicle size and zeta potential after 15 days, while the LIP-VC remained stable in its size and PDI for 30 days. After that, it is shown that VC can be used as an antioxidant and stabilizer in liposomes to increase the stability and shelf-life of vesicles.

Superoxide-anion triggers impairments of immune efficiency and stress response behaviors of Eisenia fetida earthworms.

Superoxide-hydrogen peroxide (S-HP), triggered by Val16Ala-SOD2 human polymorphism, may influence the risk of depression. Therefore, it is plausible that higher basal S-anion levels and chronic inflammatory states associated with the VV-SOD2 genotype can negatively modulate the stress response associated with resilience in various species, from primitive species to humans. To test this hypothesis, Eisenia fetida earthworms were exposed for 24 h to 30 nM rotenone, which causes mitochondrial dysfunction by generating high S-anion levels (known as the "VV-like phenotype"), and 10 µM porphyrin, a SOD2-like compound, which generates elevated HP levels (known as the "AA-like phenotype"). The results suggested that both S-anion and HP acted as signaling molecules, differentially altering the immune function and acute hydric stressful response. Although the AA-like phenotype improved the immune and stress response efficiencies, the VV-like phenotype showed a downregulated expression of the toll-like receptor (EaTLR, JX898685) and antimicrobial peptide (AMP) (AF060552) genes, which triggered the impairment of encapsulation and earthworms extracellular trap (EET) processes used by earthworms to trap and destroy microorganisms. When exposed to adverse environments and dangerous hydric stress, VV-like earthworms exhibited an impulsive behavior and failed to quickly identify and migrate to a protected environment, unlike control earthworms and AA-like earthworms. All results corroborated that the S-anion imbalance could concomitantly induce alterations in immune function and stress behavior related to earthworm survival. From a human perspective, this information may corroborate the potential specific role of superoxide anion in the modulation of the stress response, resilience, and risk of depression.

UVB photoprotective capacity of hydrogels containing dihydromyricetin nanocapsules to UV-induced DNA damage.

We evaluate the effect of cationic nanocapsules containing dihydromyricetin (DMY) flavonoid for safe topical use in photoprotection against UV-induced DNA damage. The stability was investigated for feasibility to produce hydrogels containing cationic nanocapsules of the flavonoid DMY (NC-DMY) for 90 days under three different storage conditions (4 ± 2 °C, 25 ± 2 °C, and 40 ± 2 °C), as well as evaluation of skin permeation and its cytotoxicity in skin cell lines. The physicochemical and rheological characteristics were maintained during the analysis period under the different aforementioned conditions. However, at 25 °C and 40 °C, the formulations indicated yellowish coloration and DMY content reduction. Therefore, the ideal storage condition of 4 °C was adopted. DMY remained in the stratum corneum and the uppermost layers of the skin. Regarding safety, all formulations demonstrated to be safe for topical application. NC-DMY exhibited a 50% Solar Protection Factor (SPF-DNA) against DNA damage caused by UVB radiation and demonstrated 99.9% protection against DNA lesion induction. These findings establish a promising formulation containing nanoencapsulated DMY flavonoids with a photoprotective and antioxidant potential of eliminating reactive oxygen species formed by solar radiation.

Protective effect of guarana-loaded liposomes on hemolytic activity.

Paullinia cupana var. sorbilis (Mart.) Ducke, popularly known as guarana, is one of the most promising plants in Brazilian flora and has attracted considerable interest from the scientific community owing to its numerous therapeutic activities and less side effects. Hence, using nanotechnology is a viable alternative to primarily improve the physicochemical characteristics and bioavailability of guarana. The objective of the present study was to develop, characterize, and evaluate the stability of liposomes containing guarana powder and to evaluate their antioxidant and hemolytic activity in vitro. Three different concentrations of guarana powder and two methods of liposome preparation were tested. Liposomes were developed and characterized, and their stability was analyzed by evaluating physicochemical parameters. Hemolytic activity of guarana liposomal formulation (G-Lip) was compared with that of guarana in its free form (FG) and of liposome without guarana (W-Lip). Red blood cells from rats were exposed to these different formulations dissolved in phosphate buffer solution (PBS; pH 7.4). The best stability was achieved for the formulation containing 1 mg mL-1 guarana powder produced by the reverse phase evaporation method. FG showed dose-dependent antioxidant activity, which was maintained in G-Lip. W-Lip showed high hemolytic activity in PBS at pH 7.4 possibly because of the presence of polysorbate 80, and on addition of guarana to these structures, the hemolytic process was reversed. The same protective effect was observed for FG. It is believed that the complex structure of guarana, primarily the presence of polyphenols, exerts a powerful antioxidant action, helping to protect erythrocytes.

Inhibition of filamentous fungi by ketoconazole-functionalized electrospun nanofibers.

Nanotechnology strategies have been used for delivery and controlled release of antimicrobial drugs. Electrospun nanofibers can be versatile vehicles to incorporate antimicrobials. In this work, poly-ε-caprolactone nanofibers functionalized with ketoconazole were produced by electrospinning and tested against filamentous fungi. Ketoconazole-free nanofibers were produced as controls. Functionalized nanofibers showed antifungal activity against Aspergillus flavus, A. carbonarius, A. niger, Aspergillus sp. A29, Fusarium oxysporum and Penicillium citrinum by agar diffusion test. Inhibitory zones ranging from 6 to 44mm were observed, this larger inhibition was against A. flavus. The nanofibers were incubated in different simulant solutions to evaluate the ketoconazole release, which was only detected in the solution containing 5% (v/v) Tween 20. Electron microscopy images showed the nanofibers with ketoconazole presented mean diameters of 526nm, and the degradation of the nanofiber structures could be observed by electron microscopy after incubation in simulant solution. Infrared and thermal analyses indicated that ketoconazole was dispersed without chemical interactions with the polycaprolactone matrix. These results suggest that polycaprolactone nanofibers incorporating ketoconazole may be an interesting alternative to control pathogenic fungi.

Rhizobia survival in seeds coated with polyvinyl alcohol (PVA) electrospun nanofibres.

The electrospinning technique of rhizobia immobilization in nanofibres is an innovative and promising alternative for reducing the harmful effects of environmental stress on bacteria strains in a possible inoculant nanotechnology product for use in agriculture. The use of polyvinyl alcohol (PVA) shows up as an effective polymer in cell encapsulation because of its physical characteristics, such as viscosity and power of scattering. The aim of these studies has been to evaluate the survival of rhizobia incorporated in PVA nanofibres, which were applied to soybean seed and then subjected to different storage times and exposure to fungicide. The maintenance of the symbiotic characteristics of the incorporated bacterial strains was also evaluated, noting the formation of nodules in the soybean seedlings. No significant differences in the cell survival at 0 h and after 24 h of storage were observed. After 48 h, a significant difference in the bacterial cell concentration of the seeds affixed with PVA nanofibres was observed. Exposure to the fungicide decreased the viability of the bacteria strains even when coated with the nanofibres. A larger number of nodules formed in soybean seedlings from seeds inoculated with rhizobia incorporated in PVA nanofibres than from seeds inoculated with rhizobia without PVA. Thus, the electrospinning technique is a great alternative to the usual protector inoculants because of its unprecedented capacity to control the release of bacteria.