Phytochemical analysis of carotenoid profile in Mentha piperita and Artemisia vulgaris: cytotoxicity in tumoral cells and evaluation of plasmid DNA cleavage.

Several medicinal plants have been administered to cancer patients attributed to their anticarcinogenic and chemoprotective properties, in addition to lower toxicity compared to traditional therapies. The aim was to investigate the antioxidant properties and carotenoid composition of aqueous extracts of Mentha piperita or Artemisia vulgaris which were previously found to exert beneficial effects on human health through diet. aqueous extracts exhibited potent antioxidant activity. A diversity of carotenoids was identified in these extracts using HPLC-PDA-MS/MS. Both extracts contained predominantly all-trans-lutein as the main component within this class. In order to investigate antioxidant properties, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) techniques were used. The (3-4,5 dimethylthiazol-2, 5 diphenyl tetrazolium bromide) (MTT) and Crystal Violet assays assessed cellular cytotoxicity. Assessments of presence of reactive species were carried out following exposure of oral squamous cell carcinoma cell line (SCC-4) to various aqueous extracts of M piperita or A vulgaris utilizing dichlorofluorescein diacetate (DCFH-DA) and nitric oxide (NO) assays. Exposure to these extracts induced severe cytotoxic effects, which led to investigation of the biochemical and molecular mechanisms underlying this observed effect. Data demonstrated that both solutions induced oxidative stress and DNA damage, especially at higher concentrations using agarose gel subjected to electrophoresis. It is known that exposure to excess amounts of antioxidants results in a prooxidant effect which is beneficial in cancer therapy. Further, the extracts were found to reduce viability of SCC-4 in culture, indicating that this antitumoral activity may be of therapeutic importance and requires further study.

Detection of new phytochemical compounds from Vassobia breviflora (Sendtn.) Hunz: antioxidant, cytotoxic, and antibacterial activity of the hexane extract.

Vassobia breviflora (Sendtn.) Hunz is a plant of the Solanaceae family from South America and there are no apparent studies reported on the biological activity of the hexane extract. The aim of this investigation was to conduct phytochemical analyses using ESI-TOF-MS, while antioxidant activities were evaluated by the following methods 1,1-diphenyl-2-picrylhydrazyl (DPPH) 2,2"-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical capture (ABTS), ferric reducing antioxidant power (FRAP), total antioxidant capacity (TAC), and total oxidant status (TOS). Antimicrobial activities were performed by determining the minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and antibiofilm formed. Cytotoxicity was measured by MTT and dsDNA PicoGreen tests, beyond the production of reactive oxygen species (ROS) determined by Dichlorodihydrofluorescein diacetate (DCFH-DA). The hexane extract showed the presence of 5 (choline, pantothenic acid, calystegine B, lanciphodylactone I, and 15"-cis-zeaxanthin) compounds detected. V. breviflora extract demonstrated reliable results utilizing different antioxidant methods. In antibacterial activity, V. breviflora extract exhibited inhibitory, bactericidal, and antibiofilm action in biofilm-forming bacteria. The hexane extract exhibited cytotoxicity against melanoma, lung cancer, glioblastoma, leukemia, uterine colon, and hepatocarcinoma tumor cells. In addition, all tested strains resulted in increased production of ROS. This plant extract may be considered in future as an alternative for development of new therapeutic options aimed at the treatment of diverse pathologies.

Insights of ethyl acetate fraction from Vassobia breviflora in multidrug-resistant bacteria and cancer cells: from biological to therapeutic.

Cancer and infectious diseases are among the leading causes of death in the world. Despite the diverse array of treatments available, challenges posed by resistance, side effects, high costs, and inaccessibility persist. In the Solanaceae plant family, few studies with Vassobia breviflora species relating to biological activity are known, but promising results have emerged. The phytochemicals present in the ethyl acetate fraction were obtained using ESI-MS-QTOF, and the antioxidants assays 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical capture (ABTS), plasma ferric reduction capacity (FRAP), and total antioxidant capacity (TAC). Cytotoxic activity was evaluated by MTT, Neutral Red, and lactate dehydrogenase (LDH) released. The production of reactive oxygen species, nitric oxide, and purinergic enzymes was also investigated. Antibacterial activity was measured through minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and antibiofilm activity, in addition to genotoxicity in plasmid DNA. Five major masses were identified D-glucopyranose II, allyl disulfide, γ-lactones, pharbilignoside, and one mass was not identified. V. breviflora exhibited relevant antioxidant and cytotoxic activity against the HeLa cell line and enhanced expression effect in modulation of purinergic signaling. Antibacterial activities in the assays in 7 ATCC strains and 8 multidrug-resistant clinical isolates were found. V. breviflora blocked biofilm formation in producing bacteria at the highest concentrations tested. However, there was no plasmid DNA cleavage at the concentrations tested. Data demonstrated that V. breviflora exhibited an antioxidant effect through several methods and proved to be a promising therapeutic alternative for use against tumor cells via purinergic signaling and multidrug-resistant microorganisms, presenting an anti-biofilm effect.

Eucalyptus oil nanoemulsions against eggs and larvae of Haemonchus contortus.

Haemonchus contortus is a highly pathogenic and prevalent helminth that causes many deaths in sheep herds. Anthelmintics are usually employed to overcome this issue; however, they do not guarantee immediate and lasting efficacy because of the occurrence of drug-resistant parasites. Among substances that are used in scientific studies for parasitic control, essential oils are known to have different pharmacological properties. However, they demonstrate instability owing to several factors, and therefore, nanoemulsification is considered an alternative to control the instability and degradability of these compounds. The objective of this study was to evaluate the cytotoxicity of nanoemulsions containing essential oil of Eucalyptus globulus against the blood of healthy sheep and to verify their activity against the parasite H. contortus in sheep. The results presented adequate nanotechnological characteristics (diameter 72 nm, PDI 0.2, zeta -11 mV, and acidic pH) and adequate morphology. Further, the corona effect and cytotoxic profiles of the free oil and nanoemulsion against blood cells from healthy sheep were evaluated. The tests results did not present a toxicity profile. For evaluating efficacy, we observed an important anthelmintic action of the nanoemulsion containing oil in comparison to the free oil; the results demonstrate a potential role of the nanoemulsion in the inhibition of egg hatchability and the development of larvae L1 to L3 (infective stage). Based on these results, we developed an important and potential anthelmintic alternative for the control of the parasite H. contortus.

Antioxidant and Nematocidal Effects of Several Oyster Mushroom Species of Genus Pleurotus (Agaricomycetes).

Cyathostomins are common intestinal nematodes that parasitize horses and can affect animal health and welfare. Alternative treatments are essential for sustainable parasite control, reducing the indiscriminate use of anthelmintics and their impacts on the environment, animals, and humans. Pleurotus spp. are edible medicinal mushrooms that possess several therapeutic properties, including antiparasitic and nematocidal activities. This study aimed to evaluate the antioxidant and nematocidal activity of aqueous extracts of three oyster species (P. florida, P. ostreatus, and P. djamor) on cyathostomine eggs in vitro. P. florida showed the highest activity against test organisms, inhibiting the hatching of eggs in up to 92.19% at the highest concentration (10%) of aqueous extract, with a mean lethal concentration (LC50) of 2.13%. In the aqueous extracts of P. ostreatus and P. djamor, nematocidal activity was reduced, inhibiting egg hatching by 55.46% (LC50, 7.44%) and 23.67%, respectively, at a concentration of 10%. P. florida showed higher antioxidant activity than that of the other species tested. Thus, P. florida may be considered a potential alternative for biological control in in vivo experiments with cyathostomins.

Açaí (Euterpe oleracea Mart.) presents anti-neuroinflammatory capacity in LPS-activated microglia cells.

INTRODUCTION: Neuropsychiatric diseases are responsible for one of the highest burden of morbidity and mortality worldwide. These illnesses include schizophrenia, bipolar disorder, and major depression. Individuals affected by these diseases may present mitochondrial dysfunction and oxidative stress. Additionally, patients also have increased peripheral and neural chronic inflammation. The Brazilian fruit, açaí, has been demonstrated to be a neuroprotective agent through its recovery of mitochondrial complex I activity. This extract has previously shown anti-inflammatory effects in inflammatory cells. However, there is a lack of understanding of potential anti-neuroinflammatory mechanisms, such as cell cycle involvement. OBJECTIVE: The objective of this study is to evaluate the anti-neuroinflammatory potential of an açaí extract in lipopolysaccharide-activated BV-2 microglia cells. METHODS: Açaí extract was produced and characterized through high performance liquid chromatography. Following açaí extraction and characterization, BV-2 microglia cells were activated with LPS and a dose-response curve was generated to select the most effective açaí dose to reduce cellular proliferation. This dose was then used to assess reactive oxygen species (ROS) production, double-strand DNA release, cell cycle modulation, and cytokine and caspase protein expression. RESULTS: Characterization of the açaí extract revealed 10 bioactive molecules. The extract reduced cellular proliferation, ROS production, and reduced pro-inflammatory cytokines and caspase 1 protein expression under 1 µg/mL in LPS-activated BV-2 microglia cells but had no effect on double strand DNA release. Additionally, açaí treatment caused cell cycle arrest, specifically within synthesis and G2/Mitosis phases. CONCLUSION: These results suggest that the freeze-dried hydroalcoholic açaí extract presents high anti-neuroinflammatory potential.

Phytochemical analysis and evaluation of the antioxidant and antiproliferative effects of Tucumã oil nanocapsules in breast adenocarcinoma cells (MCF-7).

In this work was to develop an inedited nanocapsule with tucumã oil (Astrocaryum vulgare). The oil presents of phytosterols (squalene and ß-sitosterol), all-trans-beta-carotene, acids oleic and palmitic. Antioxidant activity showed a good performance in DPPH and ABTS assays. The nanocapsules were prepared and demonstrated in their characterization particle size (206 ± 0.69 nm). The cytogenotoxicity evaluation was performed using the MTT, dichlorofluorescein, nitric oxide and dsDNA PicoGreen® assays. Antitumor efficacy assays in MCF-7 cells demonstrated that free oil and tucumã nanocapsules had IC50 of 130 and 50 µg/mL, respectively. Thus, previous studies of toxicity are relevant, as they generate future subsidies, aiming at the potential application of nanostructures and in addition, the promising effect of NCs of tucumã oil on the antiproliferative effect in breast adenocarcinoma cells was evidenced.

In vitro and in vivo activity of the essential oil and nanoemulsion of Cymbopogon flexuosus against Trichomonas gallinae.

OBJECTIVE: This study was done to evaluate the in vitro and in vivo effects of the essential oil (OE-CL) and nanoemulsion (N-CL) of Cymbopogon flexuosus against Trichomonas gallinae. MATERIALS AND METHODS: In vitro assays were done with 106 parasites and OE-CL and N-CL in the concentrations: 110, 220, 330, 440, 550, 660, 770 and 880 µg/ml and four controls: CN (culture medium and trophozoites), MTZ (trophozoites plus 800 µg/ml of metronidazole), TW (trophozoites plus vehicles used for solubilization of derivatives (0.01% Tween) and NB (blank nanoemulsion 880 µg/ml). The in vivo assay was done in 35 quails (Coturnix coturnix) infected experimentally 4x104 mg/kg, were divided in seven groups (n=5): A (control-healthy), B (control infected), C (control TW 0.01%), D (NB 0.88 mg/kg), E (drug MTZ 25 mg/kg, F (OE-CL at 0.55 mg/kg) and G (N-CL at 0.44 mg/kg), during 7 consecutive days. RESULTS: The in vitro test showed that the OE-CL (550 µg/ml) and N-CL (440 µg/ml) concentrations reduced the trophozoites viability in 100%. In the in vivo test, the treatment with OE-CL was efficient on the 4th treatment day and the N-CL after the 3rd day, and the MTZ in the therapeutic concentration was efficient on the 7th day. CONCLUSION: It can be observed in this study that the lemon grass has natural potential antitrichomonal activity against T. gallinae in vitro and in vivo.

Nanobiopesticides: development and inseticidal activity of nanoemulsions containing lemongrass or eucalyptus oils.

The bioinsecticides, like essential oils, are a promising alternative in pest control. However, these oils have some limitations, such as instability and low solubility. These limitations can be circumvented through nanotechnology, with the nanoemulsification of these compounds. Therefore, the objective of this study was to prepare, characterize and explore the insecticidal activity against adult flies of nanoemulsions containing essential oil of lemongrass or eucalyptus. The nanoemulsions were prepared by the high-energy method and presented droplet size smaller than 125 nm, with polydispersity index of 0.2, pH acid and spherical morphology. The insecticidal activity was evaluated by the Topical Application Method and Exposure Impregnated Paper Exposure, where it was possible to demonstrate a potential insecticidal effect of lemongrass oil in the concentrations of 10, 30 and 50 µL/mL against Musca domestica and Lucilia cuprina and the potentiation of this effect when nanoemulsified this oil against L. cuprina.

Topical hydrogel containing Achyrocline satureioides oily extract (free and nanocapsule) has anti-inflammatory effects and thereby minimizes irritant contact dermatitis.

Inflammatory dermatoses are prevalent worldwide, with impacts on the quality of life of patients and their families. The aim of this study was to determine the anti-inflammatory effects of Achyrocline satureioides oily extracts and nanocapsules on the skin using a mouse model of irritant contact dermatitis induced by croton oil, and a skin inflammation model induced by ultraviolet B (UVB) radiation. The mice were treated with 15 mg/ear oily extract (HG-OLAS) or nanocapsules (HG-NCAS) of A. satureioides incorporated into Carbopol® 940 hydrogels. We found that HG-OLAS and HG-NCAS formulations reduced ear edema in croton oil-induced lesions with maximum inhibitions of 54±7% and 74±3%, respectively. HG-OLAS and HG-NCAS formulations decreased ear edema induced by UVB radiation (0.5 J/cm2), with maximum inhibitions of 68±6% and 76±2% compared to the UVB radiation group, respectively. HG-OLAS and HG-NCAS modulated myeloperoxidase (MPO) activity after croton oil induction. Furthermore, croton oil and UVB radiation for 6 and 24 h, respectively, stimulated polymorphonuclear cells infiltration. The topical treatments reduced inflammatory processes, as shown by histological analysis. Together, the data suggest that topical application of A. satureioides oily extracts and nanocapsules produced antiedematogenic and anti-inflammatory effects. They constitute a compelling alternative for treatment of skin injuries.

Dog food production using curcumin as antioxidant: effects of intake on animal growth, health and feed conservation.

The objectives of this study were to produce dog food containing curcumin replacing synthetic antioxidants, to evaluate its beneficial effects on animal growth and health. Curcumin (100 mg/kg) was added after the extrusion process along with the other micronutrients. The final concentration of curcumin was 32.9 mg/kg. The control feed was composed of the same ingredients without curcumin. After a storage of 6 months, feed composition and pH did not differ; however, the feed with curcumin showed lower protein oxidation, lipid peroxidation and higher total antioxidant capacity. After 2 months of feed production, 12 young Beagle dogs received either curcumin-containing food (n = 6)  or the control diet (n = 6). The animals were fed twice a day using individual kennels. Blood samples were taken on d 1, 35 and 42. During the first 30 d of the study, the animals had natural infectious diseases that were controlled with anti-protozoals and antibiotics. Greater numbers of red blood cells were observed in dogs fed with curcumin (d 35 and 45), and there were greater numbers of white blood cells as a consequence of increased neutrophils on d 42. At the end of the experiment, a significant reduction in the number of lymphocytes was observed in dogs that ingested curcumin (d 42), suggesting an anti-inflammatory effect, manifested as a decrease in globulin levels. In the final 15 d of the experiment, the animals were clinical healthy. Higher serum levels of glucose, urea, triglycerides and cholesterol were observed in dogs fed with curcumin. Curcumin increased the activity of several antioxidant enzymes in addition to non-protein thiols and the total antioxidant capacity in the serum, consequently reducing levels of oxygen reactive species. Curcumin supplementation of dogs did not favour growth or weight gain. Neverthless, it was concluded that curcumin improved animal health, with emphasis on the stimulation of the antioxidant system and evidence of an anti-inflammatory effect.

Effects of curcumin and yucca extract addition in feed of broilers on microorganism control (anticoccidial and antibacterial), health, performance and meat quality.

The aim of this study was to determine whether curcumin and yucca extract addition in broiler feed improves growth, health, and meat quality, and to measure coccidiostatic and antimicrobial activity so as to enable replacement of conventional performance enhancers. We used 240 birds in four treatments: CN, basal feed with antibiotics and coccidiostatic drugs; CU, feed with 100 mg/kg of curcumin; YE, feed with 250 mg/kg of yucca extract; and CU + YE, feed with the combination of 100 mg curcumin/kg and 250 mg yucca extract/kg. A significant reduction in oocysts was observed in birds supplemented with combined additives (CU + YE) at days 37 compared to other treatments and at 42 days in relation to the CU treatment. At 42 days, the total bacterial counts for the CN and CU treatments were lower than the others. Birds fed the additive had lower numbers of leukocytes, lymphocytes, and heterophils than did those in the CN treatment. The highest levels of antioxidants in meat were observed in the treatments with the additives, together with lower levels of lipid peroxidation compared to the CN. The lowest protein oxidation was observed in the CU + YE treatment in relation to the other treatments. Lower total levels of saturated fatty acids (SFA) were observed in the CU treatment than in the CN. There were lower levels of monounsaturated fatty acids (MUFA) in the meat of birds in the YE treatment in relation to the others. Higher levels of total polyunsaturated fatty acids (PUFA) were observed in birds that consumed curcumin, individually and in combination with yucca extract. Taken together, the data suggest that curcumin and yucca extract are additives that can potentially replace conventional growth promoters; they improved bird health. Changes in the fatty acid profile of meat (increase in the percentage of omegas) are beneficial to the health of the consumer.

Benefits of nanotechnology: Dietary supplementation with nerolidol-loaded nanospheres increases survival rates, reduces bacterial loads and prevents oxidative damage in brains of Nile tilapia experimentally infected by Streptococcus agalactiae.

Rampant and uncontrolled use of antibiotics is a major concern for aquaculture; the practice foments the emergence of resistant strains of Streptococcus agalactiae, among other negative impacts. Constituents of plant essential oils such as nerolidol are being considered as replacements for synthetic drugs to support fish nutrition and health. There is evidence to suggest that nanotechnology may enhance the efficacy of natural bioactive compounds; this is a substantial advance for the development and sustainability of aquaculture. Against the backdrop of this evidence, we aimed determine whether dietary supplementation with free nerolidol and nerolidol-loaded nanospheres would exert bactericidal effects against S. agalactiae, as well as prevent S. agalactiae-induced brain oxidative damage. In Experiment I, we measured the antimicrobial properties of dietary supplementation of nerolidol and nerolidol nanosphere in terms of mortality, longevity and relative percent survival. Fish infected with S. agalactiae fed 0.5 and 1.0 mL nerolidol nanospheres kg/diet demonstrated lower mortality and higher relative percent survival than the control group, while longevity was higher in all infected plus supplementation groups. Experiment II showed significantly lower microbial loads in brains of fish infected with S. agalactiae that were fed 1.0 mL nerolidol nanospheres kg/diet than in the control group. Brain nerolidol levels were significantly higher in uninfected as well as infected fish supplemented with nerolidol nanospheres than in fish supplemented with free nerolidol. Finally, brain reactive oxygen species and lipid peroxidation levels were higher in infected fish supplemented with basal diet compared to uninfected fish and supplemented with basal diet, and the supplementation with 1.0 mL/kg nerolidol nanospheres prevented this augmentation caused by infection. These data suggest that dietary supplementation with nerolidol nanospheres (1.0 mL/kg diet) has potent bactericidal effects in terms of augmentation of fish longevity and survival, and reduction of brain microbial loads. Also, S. agalactiae-induced brain oxidative damage that contributed to disease pathogenesis, and the dietary supplementation with nerolidol nanospheres (1.0 mL/kg diet) prevented this alteration. In summary, nanotechnology is a compelling approach to enhancing the efficacy of nerolidol, giving rise to reduction of S. agalactiae loads in fish brains.

Nanospheres as a technological alternative to suppress hepatic cellular damage and impaired bioenergetics caused by nerolidol in Nile tilapia (Oreochromis niloticus).

Nerolidol is a sesquiterpene found in essential oils of several plant species. It is found commonly in human and animal diets and is approved by the US Food and Drug Administration as a flavoring agent. Nevertheless, recent studies have suggested that nerolidol has potent hepatotoxic effects. Because use of plant-based products in human and animal food has expanded considerably, it is essential to develop approaches such as nanotechnology to avoid or reduce hepatic toxic effects. Therefore, the aim of the study was to determine whether nerolidol dietary supplementation elicited hepatic damage associated with impairment of energy homeostasis, as well as whether supplementation with nerolidol-loaded in nanospheres prevented hepatotoxic effects in Nile tilapia (Oreochromis niloticus). Nile tilapia were divided into five groups (A-E, n = 10 per group) with four replicates each, as follows: group A received basal feed (without supplementation); group B received feed containing 0.5 mL free nerolidol/kg; group C received feed containing 1.0 mL free nerolidol/kg; group D received feed containing 0.5 mL nanospheres nerolidol/kg; and group E received feed containing 1.0 mL nanospheres nerolidol/kg. All groups received experimental feed once a day (10% total biomass) at 2 p.m. for 60 consecutive days. Hepatic liver weight and relative liver weight were significantly lower in fish fed 1.0 mL free nerolidol/kg feed than in fish given basal diet (control group). Hepatic pyruvate kinase (1.0 mL free nerolidol/kg) and adenylate kinase (0.5 and 1.0 mL free nerolidol/kg) activities were significantly lower than in the control group, while hepatic reactive oxygen species and lipid damage levels were significantly higher. Finally, the comet assay revealed significant increases in the frequency of damage and the damage index in fish given 0.5 and 1.0 mL free nerolidol/kg in a dose-dependent manner. Nerolidol-loaded in nanospheres prevented all alterations elicited by free nerolidol. Based on these data, we concluded that dietary supplementation with free nerolidol elicited severe impairment of hepatic bioenergetics homeostasis that appeared to be mediated by excessive ROS production and lipid damage, contributing to a genotoxic effect. Dietary supplementation with nerolidol-loaded in nanospheres did not elicit hepatic damage, and therefore, should be considered as a replacement so as to limit toxicity, permitting its continued use as a dietary supplement.

Low-dose curcumin-loaded Eudragit L-100-nanocapsules in the diet of dairy sheep increases antioxidant levels and reduces lipid peroxidation in milk.

The main objective of this study was to evaluate whether the addition of curcumin-loaded nanocapsules (prepared and characterized) in the diets of dairy sheep improved milk quality. The nanocapsules were prepared using two polymers: poly-ε-caprolactone (PCL) and Eudragit L-100. The nanocapsules contained 0.25 mg/ml (Nano-Eudragit L-100) and 2 mg/ml (Nano-PCL) of curcumin. Dairy sheep were divided into four groups: A (control), B (30 mg free curcumin/kg concentrate), C (3 mg Nano-PCL/kg concentrate), and D (3 mg Nano-Eudragit/kg concentrate). We observed that the number of total leukocytes and serum globulin levels were lower in Group D than in the control (Group A) (p < 0.05). Antioxidant capacity against peroxyl radicals (ACAP) and catalase enzymes was elevated in Group D, with consequently reduced lipid peroxidation (LPO; p < 0.05). In milk, there were no differences in production and composition between groups during the experimental period (p > 0.05); however, ACAP increased and LPO decreased in milk. PRACTICAL APPLICATIONS: Curcumin is a functional molecule with potent antioxidant, anti-inflammatory, and antimicrobial actions, used frequently and with medical indications in human food. Free curcumin in sheep diets improves milk quality and increases its shelf life. This study showed that curcumin nanocapsules produced from the Eudragit L-100 polymer potentiated the anti-inflammatory and antioxidant actions of dairy sheep when used in the diet daily, at doses 10 times lower than that of free curcumin. These positive effects were reflected in higher total antioxidant capacity and lower lipid peroxidation in milk in sheep-fed curcumin-loaded Eudragit L-100 nanocapsules, generating desirable milk properties. In practice, the use of nanotechnology enhances the beneficial effects of curcumin in milk, possibly creating a nutraceutical food desirable to consumers.

Effect of free and nano-encapsulated curcumin on treatment and energetic metabolism of gerbils infected by Listeria monocytogenes.

Bacterial infections require special care since the indiscriminate use of antibiotics to treat them has been linked to the emergence of resistant strains. In this sense, phytoterapeutic alternatives such as curcumin and its nanocapsules have emerged as a promising supplement in optimizing availability of bioactives and reducing the development of antimicrobial resistance. Thus, the aim of this study was to verify the effects of pure and nanoencapsulated curcumin in the treatment of experimental listeriosis in gerbils regarding many aspects including antibacterial effect, antioxidant mechanisms involved and the energetic metabolism. Four groups were used containing 6 animals each: T0 (control), T1 (infected), T2 (infected and treated with free curcumin - dose of 30 mg/kg/day) and T3 (infected and treated with nanocapsules containing curcumin - a dose of 3 mg/kg/day). Treated animals received curcumin for 6 consecutive days starting 24 h after Listeria monocytogenes infection. All animals were euthanized on the 12th day after L. monocytogenes infection. Quantitative polymerase chain reaction (qPCR) identified L. monocytogenes DNA in the spleens of all animals of the T1 group, as well as T2 (2 out of 6) and T3 (5 out of 6). The weight of the spleens confirmed the infection, since it was larger in the T1 group, differing statistically from T0, and similarly to T2 and T3. Hepatic histopathological examination showed mild infiltration of neutrophils and macrophages, except for the T3 group (only 1/6). In the liver, the pyruvate kinase activity was higher in T1 and T2 compared to T0 and T3. The adenylate kinase activity did not differ between groups. The Na+/K+ATPase activity was lower in T1 group compared to T0 and T3. Lipoperoxidation was lower in the T3 group compared to groups T0, T1 and T2. The antioxidant capacity against peroxyl radicals was higher in T1, T2 and T3 groups compared to T0. In conclusion, free curcumin showed potent antibacterial effects; however, the nanoencapsulated form was able to minimize the effects caused by L. monocytogenes regarding tissue injury, changes on enzymes of the energetic metabolism, in addition to an antioxidant effect against lipoperoxidation.

Hydrogel containing silibinin nanocapsules presents effective anti-inflammatory action in a model of irritant contact dermatitis in mice.

The current study developed an innovative Pemulen® TR2 hydrogel containing silibinin-loaded pomegranate oil-based nanocapsules (HP-NC SB) intending cutaneous application. The formulation anti-inflammatory activity in an in vivo model and biometric studies on the skin of healthy volunteers were also performed. The nanocapsules were prepared using the interfacial deposition of preformed polymer technique and the hydrogels were obtained by thickening of nanocapsules suspension with Pemulen® TR2. Formulations with free compound, vehicle and blank nanocapsules were also produced. The hydrogels were evaluated concerning pH, silibinin content, particle size, spreadability profile, rheology, in vitro drug release, cutaneous permeation, bioadhesive potential and cutaneous biometry evaluation. Furthermore, a model of contact dermatitis croton oil-induced in mice was performed to evaluate the hydrogels anti-inflammatory potential. The formulations presented adequate characteristics for skin administration: particle within nanometric size, pH values in the acid range, silibinin content close theoretical values (1 mg/g) and non-Newtonian pseudoplastic behavior. Nano-based hydrogels showed high bioadhesive properties, increased silibinin in vitro release profile and its retention in the stratum corneum. The best anti-inflammatory effect was exhibited by HP-NC SB, which reduced both ear edema and inflammatory cells infiltration in comparison to the induced group. Furthermore, cutaneous biometric evaluation showed that formulations containing free or nanoencapsulated silibinin caused no modification in normal skin conditions (pH, tissue hydration, transepidermal water loss and erythema). In summary, the results demonstrated that the Pemulen® TR2 hydrogel containing NC SB was successfully developed, indicating its potential as an alternative treatment for irritant contact dermatitis.

Low Dose of Nanocapsules Containing Eucalyptus Oil Has Beneficial Repellent Effect Against Horn Fly (Diptera: Muscidae).

For the first time, the repellent and insecticidal effects of eucalypt essential oil (Eucalyptus globulus) in its free form and associated with different nanostructures (nanoemulsion and nanocapsules) were investigated against Musca domestica (Diptera: Muscidae) and Haemotobia irritans (Diptera: Muscidae) flies. Specimens of M. domestica were collected, separated into groups (n = 10), treated with aspersion of essential oil of eucalypt in its free (1, 5, and 10%) and nanostructured (nanoemulsion or nanoencapsulated at 1, 3, and 5%, respectively) forms. The determination of the insecticidal effect was performed by counting the number of dead flies at intervals of 30, 60, 120, 360 and 750 min after oil aspersion. E. globulus essential oil showed insecticidal effect against M. domestica at concentrations of 1 and 5%. Eucalypt essential oil (10%) caused 100% mortality of M. domestica after 750 min of the treatment. Nanocapsules (3 and 5%) showed efficacy by reducing the number of flies. On the other hand, nanoemulsion did not show insecticidal effect. The repellent action of E. globulus concentrations of 5% was tested against H. irritans on naturally infested cows. The repellent action against H. irritans was verified soon after pulverization. After 24 h, a significant reduction on horn flies' population (83.33 and 66.66%) was observed using free and nanoencapsulated forms tested, respectively. E. globulus essential oil demonstrated insecticidal and repellent effects against M. domestica and H. irritans flies.

Protective effect of nerolidol-loaded in nanospheres against cerebral damage caused by Trypanosoma evansi.

Trypanosoma evansi is a zoonotic parasite associated with high animal mortality that has gained importance due to its capacity to infect humans. Recently, some evidences have demonstrated that T. evansi infection causes severe genotoxic and cytotoxic damage in brain cells, contributing to the pathogenesis and clinical signs of the disease. In this sense, the aim of this study was to evaluate whether nerolidol-loaded in nanospheres, a natural compound with trypanocidal and neuroprotective effects, is able to protect the brain tissue from the cytotoxic and genotoxic effects found during T. evansi infections. Trypanosoma evansi induced brain genotoxic effects through increased damage index (DI) and frequency of damage (FD) when compared to the control group. Moreover, T. evansi induced cytotoxic effects through the reduction of brain cell viability compared to the control group. The metabolites of nitric oxide (NO x ) increased in infected animals compared to the control group. The treatment with nerolidol-loaded in nanospheres prevented the increase on brain DI, FD, and NO x levels, as well as the reduction on cell viability. Based on these evidences, these results confirm that T. evansi induces genotoxic and cytotoxic damage mediated by the upregulation of NO x levels. The most important finding is that nerolidol-loaded in nanospheres was able to prevent DNA damage and cell mortality through the modulation of brain NO x levels. In summary, this treatment can be considered an interesting approach to prevent T. evansi brain damage due its anti-inflammatory property.

Nanoemulsions containing Cymbopogon flexuosus essential oil: Development, characterization, stability study and evaluation of antimicrobial and antibiofilm activities.

The increase of microbial resistance generates the search for new substances with antimicrobial potential. The essential oil of Cymbopogon flexuosus (Lemongrass) stands out in the literature for its antimicrobial, insecticide and antioxidant properties, but it has high volatilization and low stability, and the nanoencapsulation of this oil could be an alternative to overcome these limitations. Thus, the objective of this study was to develop, for the first time, nanoemulsions containing the essential oil of C. flexuosus, through a method that does not use organic solvent and with temperature control to avoid the volatilization of the oil, characterize and evaluate of stability and the antimicrobial and antibiofilm activities of these nanoemulsions. Nanoemulsions presented adequate physicochemical characteristics (average size less than 200 nm, polydispersity index less than 0.3, negative zeta potential and acid pH) which were maintained during 90 days of storage, and the nanoencapsulation of the C. flexuosus oil enhanced its therapeutic efficacy against the microorganisms evaluated in this study compared to the free oil. These results are very promising because among the microorganisms that the nanoemulsion containing C. flexuosus was able to inhibit the formation of biofilm are the bacteria Pseudomonas aeruginosa and Staphylococcus aureus, which were recently listed by the World Health Organization as priority pathogens for development of new antibiotics.