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In this work, we report, for the first time, the magnetite-functionalization and biological evaluation of eugenol by the co-precipitation method employed only Fe2+ under mild conditions and control from the amount of the incorporated magnetite. Magnetic nanoparticles were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared (FTIR), and hydrodynamic size distribution (Zetasizer). SEM images showed that EUG·Fe3O4 is 200 nm in size and similar in shape to a nanoflower. The FTIR spectrum confirmed the presence of characteristic EUG and Fe3O4 bands in the EUG·Fe3O4 sample, while the XRD analysis showed that the magnetite functionalization with eugenol slightly affected the Fe3O4 crystal structure. The in vitro safety profile and cytotoxicity of free eugenol, magnetite pristine, EUG·Fe3O4 1:1, EUG·Fe3O4 1:5, and EUG·Fe3O4 1:10 was investigated using human cell lines (keratinocytes and melanoma). The results demonstrate the high biocompatibility of EUG·Fe3O4 in HaCat cells and the greater specificity for the A375 cell line. Furthermore, the magnetite-functionalized with eugenol decreased the toxic effects of free eugenol on healthy cells. Antibacterial tests were performed in different bacterial strains. The experimental data showed that among the magnetic compounds, the microorganisms were only sensitive to treatment with EUG·Fe3O4 1:1. Regarding the antibiofilm activity assay, it can be observed that only the EUG·Fe3O4 caused a significant decrease in biomass when compared to the positive control. Finally, it can be concluded that EUG·Fe3O4 proves to be a potential candidate for future studies for drug delivery of cancer and bacterial infections treatments.

To present a possible new alternative for wound treatment, this work evaluated the biological safety and therapeutic efficacy of graphene oxide (GO) and reduced graphene oxide (rGO) nanoparticles (NPs). First, the nanostructures were studied in silico and showed to be able to inhibit the production of some pro-inflammatory cytokines and stimulate the production of the anti-inflammatory cytokine IL-10, especially rGO. The results of the morphological and structural characterization of GO NPs synthesized from the Hummers method and reduced by ascorbic acid, were consistent with the literature, confirming their achievement. In the broth microdilution assay, GO and rGO showed antimicrobial activity against the clinical isolate of Streptococcus agalactiae (S. agalactiae) at a minimum inhibitory concentration (MIC) of 625 µg/mL for GO and 312.5 µg/mL for rGO. In addition, the nanostructure of rGO was able to inhibit, in subinhibitory concentration, the formation of S. agalactiae biofilm by up to 77% when compared to the positive control. Both NPs, in all tested concentrations, did not cause hemolysis, and alterations in coagulation in vitro assays. However, in the safety tests, it was evidenced that only the MIC of 312, µg/mL for rGO was biologically safe and presented anti-inflammatory and healing behavior in vitro. In general, the present work confirmed rGO's potential in the treatment of chronic wounds, since in silico showed anti-inflammatory behavior and in vitro showed therapeutic efficacy at low concentrations, prevented biofilm formation, and showed no significant toxic effects.

Biofilms are responsible for up to 80% of antimicrobial-resistant nosocomial infections. Most of these infections are associated with medical devices such as urinary catheters, and in this context, it is estimated that 90-100% of patients who undergo long-term catheterization develop infections. Proteus mirabilis, the most prevalent microorganism, is responsible for 20-45% of these infections. Thus, this study aimed to evaluate, for the first time, the antimicrobial and antibiofilm effects of cationic porphyrins on P. mirabilis. Neutral porphyrins 3-H

In this study, B. cereus was detected in dialysis fluids within international parameters (ultrapure – maximum limit of 0.1 CFU/mL for heterotrophic bacteria count) by analyzing the pellet obtained through the centrifugation method. We also investigated the ability of the B. cereus isolate to form a biofilm at different temperatures, the production of virulence factors, and the susceptibility to commercial antimicrobial agents. This study demonstrated a high ability of B. cereus to persist in the hemodialysis system, which can be explained by its broad ability to produce a biofilm at 25 °C, its relevant production of virulence factors, such as β-hemolysin, lecithinase and cereulide, and its important resistance pattern to antimicrobial drugs. In conclusion, these new findings expand the understanding that this microorganism should not be neglected and new methods for tracking it should be considered.

Microbial infections caused by sessile microorganisms are known to be a more challenging issue than infections caused by the same microorganisms in the planktonic state. Pseudomonas aeruginosa is an opportunistic pathogen and biofilm-forming agent. This species presents intense cellular communication mediated by signaling molecules. This process is known as quorum sensing (QS) and induces the transcription of specific genes that favors cell density growth and three-dimensional bacterial grouping. In this context, the discovery of compounds capable of inhibiting the action of the QS signaling molecules seems to be a promising strategy against biofilms. This work aimed to evaluate the anti-biofilm action and the in vitro safety profile of a sulfamethoxazole-Ag complex. The results obtained indicate potential anti-biofilm activity through QS inhibition. In silico tests showed that the compound acts on the las and pqs systems, which are the main regulators of biofilm formation in P. aeruginosa. Additionally, the molecule proved to be safe for human peripheral blood mononuclear cells.

Ethnopharmacological relevance: Tucumã (Astrocaryum vulgare)is a fruit native to the Amazon region. Extracts from the peel and pulp are thought of as promising treatments for bacterial infections. The primary constituents of Tucumã oil and butter possess unsaturated carbon chains that are susceptible to oxidation by light or heat. The oils have high volatility and low aqueous solubility that limits their use without a vehicle. Nanotechnology refers to techniques to solve these problems. Nanostructured lipid carriers (NLC), for example, protect fixed oils degradation by heat or light, as well as from oxidation and evaporation, ensuring greater stability and function, thereby prolonging the useful life of the final product. Study objectives: The objective of this study was to evaluate the hemolytic, cytotoxic, antimicrobial and antibiofilm properties of an NLC containing Tucumã butter and oil soasto improve the solubility and photosensitivity of the compounds, generating better pharmacological efficacy. Materials and methods: The NLC was assessed for stability for 60 days. The cytotoxicity of nanoparticles in peripheral blood mononucleated cells was determined in culture using assays for cell viability, DNA damage, oxidative metabolism and damage to human erythrocytes. Antimicrobial activity was determined using the broth microdilution technique and antibiofilm activity according to standardized protocols. Results: The Tucumã NLC remained stable throughout the evaluated period, with pH between 5.22–5.35, monodisperse distribution (PDI

This manuscript presents the cytotoxicity, antimicrobial activity, antibiofilm preliminary properties, and associated therapy with commercial drugs using water-soluble tetra-cationic porphyrins against Pseudomonas aeruginosa. Two commercial tetra-cationic porphyrins were tested against a standard strain of P. aeruginosa 01 (PA01) in antibacterial activity assays under dark conditions and irradiated with white light for 120 min. Porphyrin 4-H

Herein, we present a new variation for an eco-friendly methodology for the synthesis of chalcogenoester in good-to-excellent yields in a short time, with an easy work-up/purification step, and in a greenest methodology, affording the minimum generation of solid and liquid waste, in comparison to that described in the literature. Additionally, some selected compounds were evaluated as antimicrobial agents, showing moderate activity against a variety of microorganisms including the K. pneumoniae, P. aeruginosa and also some selected fish pathogenic bacteria.

A utilizacao inadequada de farmacos antimicrobianos desencadeou o aumento de cepas resistentes aos tratamentos. Este cenario se torna mais preocupante quando os microrganismos se apresentam na forma de biofilme. Os biofilmes sao estruturas complexas de aglomerados microbianos, protegidos por uma camada de exopolissacarideos (EPS) que impede a penetracao dos farmacos, dificultando ainda mais o tratamento das infecoes. Com isso, o emprego de plantas medicinais como alternativa de tratamento antimicrobiano esta crescendo no mercado farmaceutico, pois estas possuem metabolitos secundarios, que sao os principais responsaveis pelas atividades biologicas das plantas. Tendo em vista os metabolitos secundarios como fonte de pesquisa, em nosso estudo, destacamos a classe das cumarinas, que sao compostos fenolicos presentes em uma vasta diversidade de vegetais e possui relatos de atividades antimicrobiana, antioxidante e antiviral. Diante disso, este estudo tem por objetivo avaliar a atividade antibiofilme da cumarina frente a biofilme formado por P. aeruginosa (PA01). A atividade antibiofilme foi avaliada por tecnica semi-quantitativa em placas de 96 pocos. O biofilme foi revelado utilizando solucao de cristal violeta e o biofilme solubilizado foi submetido a leitura em 570nm. Alem disso, foi realizada a contagem de celulas sesseis. Os resultados observados demonstraram que a cumarina e capaz de inibir a formacao do biofilme em ate 78% e reduzir a contagem de colonias sesseis. Haja vista esses resultados, a cumarina apresenta significativamente atividade antibiofilme, com isso sugerimos que esta substância pode ser utilizada como alternativa de tratamento para infeccoes que envolvem a formacao de biofilmes.

Pullulan (PUL) films containing pomegranate seed oil and Eudragit

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.

Brazilian fish farming goes together with the emergence of numerous bacterial diseases, with Aeromonas hydrophila being the main bacterial pathogen. As a consequence, antimicrobials are excessively used. Considering that antimicrobials are relatively stable and nonbiodegradable, medicinal plants and their phytochemicals have been used as alternative treatments of bacteriosis in fish farming. Limonene is a monoterpene available in two enantiomers: R-(+)-limonene and S-(-)-limonene. This study analyzed the antibacterial activity of the phytochemicals S-(-)-limonene and R-(+)-limonene against some bacteria isolated from silver catfish (Rhamdia quelen). Furthermore, by means of spectrophotometry and atomic force microscopy, we also investigated the combination therapy of phytochemicals with antimicrobials and their activity in terms of inhibiting biofilm formation. Six clinical isolates and a standard strain were selected for antimicrobial activity testing. Biofilm formation was tested in 96-well plates and nylon cubes. The most sensitive of the strains tested was the A. hydrophila strain (MF 372510). S-(-)-limonene and R-(+)-limonene had high minimum inhibitory concentrations, however, they strongly inhibited A. hydrophila biofilm formation. R-(+)-limonene and S-(-)-limonene had an additive effect when combined with florfenicol and an antagonistic effect with oxytetracycline. In general, the phytochemicals tested showed strong antibiofilm activity against A. hydrophila, and when in combination therapy with florfenicol, they showed an additive effect against the treatment of A. hydrophila.

The cubiu (Solanum sessiliflorum) is a tropical fruit native to the Amazon region and widely used in medicine and cosmetics, despite the lack of research regarding the actual safety and effectiveness of its use for these purposes. This study aimed to evaluated the phytochemical characterization, pharmacological properties (skin wound healing, antimicrobial and anti-inflammatory properties), and toxicity of cubiu extract. The cubiu antimicrobial capacity was determined against strains of Aeromonas caviae, Pseudomonas aeruginosa, and Sphingomonas paucimobilis. Additionally, cubiu toxicity (hemolysis, coagulation, cell viability, and genotoxicity tests), antioxidant activity (reactive oxygen species total levels), scratch assay (in vitro skin wound healing), and anti-inflammatory activity against phytohemagglutinin and in the scratch assay (Interleukin 1, interleukin 6, interleukin 10, tumor necrosis factor-alpha, and interferon-gamma levels), were evaluated. Human fibroblast cells were used to evaluate skin wound healing, and human peripheral blood mononuclear cells were used for the other assessments. Our findings showed that the cubiu extract is rich in phenolic compounds, the major compound being 5-caffeoylquinic acid. In addition, was effective against the three bacterial strains tested and inhibited and destroyed the biofilm formed by Pseudomonas aeruginosa. The cubiu extract also was no toxicity, maintained the hemocompatibility parameters in the biological range, improved cell viability, decreased reactive oxygen species total levels and pro-inflammatory cytokine levels, increased anti-inflammatory cytokine levels, and accelerated the wound healing process. In conclusion, this is the first research to prove that cubiu is an important compound for use in the skin diseases, promoting skin wound healing, antimicrobial and anti-inflammatory activities.

Microbial infections caused by sessile microorganisms are known to be a more challenging issue than infections caused by the same microorganisms in the planktonic state.

The protein A13-2 was obtained from Bacillus thuringiensis strains isolated from the Papaloapan watershed region (Oaxaca, Mexico). The cytotoxic activity of parasporal inclusions was studied against breast cancer cell line (MCF-7) and normal cell (human peripheral blood mononuclear cells). The MTT, the formation of reactive species, nitric oxide, free cell DNA, and the type of death cellular were assessed. The protein A13-2 shows the highest cytotoxic activity against MCF-7 (13% cell viability at 6 µg/mL), the extracellular DNA increases, and it shows no stress for reactive species or nitric oxide. Besides, the A13-2 parasporin shows no toxicity to peripheral blood mononuclear cells, and it does not generate changes in nitric oxide levels or free cell DNA. Due to that, the cytotoxic effect of A13-2 was specific for MCF-7, and it does not affect normal cells. According to microscopy and flow cytometry, A13-2 parasporin leads to the death of MCF-7 cells by late apoptosis together with necrosis and without allowing the triggering of the survival mechanisms. When analyzed together, our results show for the first time that the A13-2 protein isolated from Mexican strains of B. thuringiensis preferentially kills MCF- 7 (cancer cells) over HEK 293 and PBMC cell lines (normal cells), thus representing a promising alternative for the treatment of cancer breast.

Neurodegenerative diseases are multifactorial debilitating disorders of the nervous system affecting approximately 30 million individuals worldwide. Mitochondrial dysfunction and oxidative stress have also been implicated in causing neurodegeneration. As life expectancy is increasing, neurodegenerative disorders are becoming a major social issue. None of the drugs currently available for treatment are capable of healing the patient. This means that new molecules should be explored. Plants have been used for treatment of countless medical conditions and extensive research is being carried out on species of the Myrtaceae family, widely used in traditional medicine. To date, Myrciaria plinioides D. Legrand has not been studied for its therapeutic use. To evaluate the neuroprotective effect of aqueous and ethanol extracts of this plant, we investigated the protective effects in human neuroblastoma cells (SH-SY5Y). High-performance liquid chromatography fingerprinting of extracts revealed the presence of phenolic compounds and flavonoids. Extracts showed antioxidant activity in the ORAC, DPPH, FRAP and GAE methods. Ethanol extract presented a strong inhibitory activity toward p38 and JNK3 MAPKs and AChE activity and also toward TNF-α release in human whole blood. None of the extracts significantly affected cell viability; the ethanol extract, however, reversed 6-OHDA-induced toxicity. Particularly the ethanol extract suggests neuroprotective effects by preventing membrane depolarization and by significantly decreasing H2O2 production and caspase-3 activity. The present results indicate that the ethanol extract protects SH-SY5Y cells against oxidative damage and apoptosis, as shown by the antioxidative activity of the extract as well as by the inhibition of important proteins such as caspase-3, p38 and JNK3 and the cytokine TNF-α. info:eu-repo/semantics/publishedVersion

The pathophysiological mechanisms associated with immune responses to the opportunistic bacterial pathogen Providencia rettgeri in fish species are unknown. The aim of this study was to evaluate whether purinergic signaling generates anti-inflammatory or pro-inflammatory profiles in lymphoid immune organs of the Nile tilapia (Oreochromis niloticus) experimentally infected by P. rettgeri. Adenosine triphosphate (ATP) and adenosine diphosphate (ADP) hydrolysis activities in the spleen and triphosphate diphosphohydrolase (NTPDase) in the head kidney decreased in experimentally P. rettgeri-infected Nile tilapia on days seven and fourteen post-infection (PI) compared to the control group. At these same time points, splenic and head kidney adenosine deaminase (ADA) activity, and levels of ATP, interleukin-15 (IL-15) and metabolites of nitric oxide (NOx) increased in infected animals compared to their respective control groups; this was also observed for serum ATP levels. No significant difference was observed between groups regarding 5′-nucleotidase activity. These results suggest that purinergic signaling in lymphoid immune organs of Nile tilapia exerts a pro-inflammatory profile in response to P. rettgeri infection. This response contributes to the inhibition of immune and inflammatory responses by reduction of ATP hydrolysis and augmentation of ATP in the extracellular environment as well as augmentation in adenosine deamination and its possible reduction in the extracellular environment, leading to a self-sustained pro-inflammatory cycle, corroborated by excessive content of ATP (extracellular milieu), IL-15 and NOx levels. Purinergic signaling may thereby contribute to the pathophysiology of P. rettgeri infection.

Bacterial diseases are one of the major problems in freshwater fish culture and have been linked to significant losses and high mortality rate. In this study, Nile tilapia Oreochromis niloticus was infected by Providencia rettgeri to evaluate the oxidative stress and antioxidant responses in the fish tissues. Juvenile Nile tilapia was divided into two groups, as follow: control (uninfected) and experimentally infected with 100 μL of P. rettgeri suspension containing 2.4 × 107 viable cells/fish, and the liver and kidney tissues were collected on days 7 and 14 post-infection (PI). Liver and kidney ROS and lipid peroxidation levels were high in infected fish on day 14 PI compared to control group, while superoxide dismutase activity was lower in liver (days 7 and 14 PI) and kidney (day 14 PI) compared to their respective control groups. Liver and kidney antioxidant capacity against peroxyl radicals, non-proteic, and proteic thiols levels was lower in infected tilapia on day 14 PI compared to control group. Based on these results, P. rettgeri infection may elicit oxidative damage via increased ROS production, decreased ROS elimination and inhibits enzymatic and non-enzymatic antioxidant defense systems; which may contribute directly to disease pathophysiology of infected animals.

Grape pomace is a by-product of the winemaking process, rich in bioactive compounds such as resveratrol and has antioxidant and antimicrobial properties. Recent evidence has demonstrated the protective effects of grape against impairment of the phosphoryl transfer network, a system linked to energetic homeostasis of high-energy phosphoryl transfer, to maintain energetic balance. Also, Pseudomonas aeruginosa infection in fish has been related to severe impairment of the branchial phosphoryl transfer network via inhibition of creatine kinase (CK), adenylate kinase (AK) and pyruvate kinase (PK) activities. Thus, the aim of this study was to evaluate whether dietary supplementation with grape pomace flour (GPF) is able to reduce or prevent the impairment of cellular energetic homeostasis in grass carp (Ctenopharyngodon idella) experimentally infected by P. aeruginosa. Branchial CK (cytosolic and mitochondrial), AK, and PK activities were lower in infected animals on day 15 post-infection (PI) compared to uninfected fish, while lactate dehydrogenase (LDH) activity was higher. Branchial reactive oxygen species (ROS), thiobarbituric acid reactive substances (TBARS), lipid oxidation (LOOH) and protein carbonylation levels were higher, while the level of non-proteic (NPSH) and proteic (PSH) thiols was lower in infected fish on day 15 PI, as compared to uninfected fish. The use of 300 mg of GPF/kg of feed was able to ameliorate the activities of CK, AK, PK, and LDH as compared to infected fish, but their activities remained different from the control group. On the other hand, dietary supplementation with 300 mg of GPF/kg of feed was able to prevent all alterations linked to oxidative damage elicited by disease. Based on this evidence, inhibition of the phosphoryl transfer network may contribute to the impairment of intracellular energetic communication between cellular ATP synthesis and consumption and is mediated by oxidative damage in the gills of fish infected by P. aeruginosa. The use of 300 mg of GPF/kg of feed exerted protective effects on branchial energetic metabolism linked to ATP metabolism by reducing the impairment in cellular energetic homeostasis; its effects can be mediated by the prevention of SH group oxidation.

Pseudomonas aeruginosa is a Gram-negative opportunistic bacterial pathogen in aquaculture systems being associated to extensive liver damage caused by oxidative stress in both marine and freshwater fish. Dietary supplementation with natural antioxidants is considered a rational strategy to prevent hepatic diseases involved with oxidative stress. Bio-residues resulting from the wine industry, such as grape pomace, are potential sources of bioactive phenolic compounds that can be applied as supplement for animal production. Thus, the aim of this study was to evaluate whether dietary supplementation with grape pomace flour (GPF) was able to prevent or reduce the hepatic oxidative damage of grass carp, Ctenopharyngodon idella, experimentally infected by P. aeruginosa. Hepatic reactive oxygen species (ROS), metabolites of nitric oxide (NOx), thiobarbituric acid reactive substances, and protein carbonylation levels were higher in fish experimentally infected by P. aeruginosa compared to the control group. Hepatic superoxide dismutase and catalase activities and antioxidant capacity against peroxyl radical levels were also higher in fish experimentally infected by P. aeruginosa compared to the control group. Dietary supplementation with 300 mg/kg GPF prevented all alterations elicited by P. aeruginosa, with the exception of protein carbonylation levels. The dietary supplementation with 150 mg/kg GPF was not able to avoid alteration of the analyzed variables, being results similar to those infected (positive control). Based on these results, dietary supplementation with 300 mg/kg GPF prevented P. aeruginosa-induced liver damage in grass carp, and this protective effect occurred through prevention on excessive ROS and NOx production, as well as via prevention of lipid damage. Moreover, 300 mg/kg GPF exerted its hepatoprotective effects by improving enzymatic and non-enzymatic antioxidant defense system. In summary, this supplementation can be an interesting approach to prevent P. aeruginosa-induced liver damage.

It is becoming evident that bacterial infectious diseases negatively affect gill energy metabolism, mainly because it causes alterations of enzymes that belong to the phosphotransfer network, such as creatine kinase (CK), adenylate kinase (AK) and pyruvate kinase (PK). Thus, the aim of this study was to evaluate whether the phosphotransfer network could be one pathway involved in the bioenergetic imbalance during Providencia rettgeri infection, as well as to investigate the bacteria's influence on adenosine triphosphate (ATP)-dependent enzymes. In this study, branchial cytosolic and mitochondrial CK activity decreased in infected animals on day 14 post-infection (PI) when compared to its respective control group. Moreover, branchial sodium-potassium pump (Na+/K+-ATPase) and proton-pump (H+-ATPase) activities decreased in infected animals on day 14 PI compared to the control. Finally, branchial thiobarbituric acid reactive substance levels increased in infected animals on day 14 PI, while non-protein thiol (NPSH) levels decreased. No significant difference was observed between groups regarding AK and PK activities. In summary, the impairment of CK activity elicited by P. rettgeri caused an impairment in branchial energetic homeostasis, reducing ATP availability in the gills. Moreover, inhibition of CK activity provokes an impairment of ATP-dependent enzymes, such as Na+/K+-ATPase and H+-ATPase, which can be mediated by lipid oxidative damage and reduction of NPSH content.

The spleen is a secondary lymphatic organ that has an important role in fish immune and inflammatory responses. Reports have suggested a role of purinergic signaling in immune and inflammatory responses during bacterial infection; however, the role of this pathway during Pseudomonas aeruginosa infection remains unknown. The aim of this study was to investigate the involvement of purinergic signaling in the pathogenesis of P. aeruginosa infection of grass carp, Ctenopharyngodon idella. In addition, grape pomace flour (GPF), a bio-residue of the wine industry, was assessed as a dietary supplement and whether it was able to modulate purinergic signaling and improve the immune and inflammatory responses of C. idella, experimentally infected with P. aeruginosa. Nucleoside Triphosphate diphosphohydrolase (NTPDase) activity was downregulated in the serum and spleen of fish experimentally infected with P. aeruginosa, compared to the control group. By contrast, serum and splenic adenosine deaminase (ADA) activity was stimulated in fish experimentally infected with P. aeruginosa compared to the control group. Dietary supplementation with GPF was able to prevent all alterations induced by infection. Serum and splenic metabolites of nitric oxide (NOx) were higher in fish experimentally infected with P. aeruginosa compared to the control group, and GPF supplementation was able to prevent this alteration. Serum and splenic myeloperoxidase and catalase activities were also higher in fish experimentally infected with P. aeruginosa compared to the control group, and again, the use of GPF was able to prevent these alterations. Based on these metabolic changes, purinergic signaling contributed to the pro-inflammatory profile of immune lymphatic organs during P. aeruginosa infection and to disease pathophysiology. Dietary supplementation with GPF modulated the inflammatory response by altering serum and splenic purinergic signaling. GPF may be means of improving immune and inflammatory responses resulting from P. aeruginosa infection of grass carp.

Objective: Evaluate methylene blue (MB) formulations containing oxygen carrier at different pHs in antimicrobial photodynamic therapy (aPDT). Material and Methods: Biofilms of Pseudomonas aeruginosa PA01 formed over acrylics specimens during five days were treated with aPDT using different formulations: MB/pH 7.4; MB/pH 5.6; MB/carrier pH 7.4; MB/carrier pH 5.6. Biofilms not exposed to treatment were used as a control. Blind examiner for the experimental groups performed the counting of colonies per ml suspension (CFU/ml). Two-way ANOVA was used to determine the effect of factors solvent (carrier vs water) and pH (7.4 vs 5.6). One-way ANOVA and post-hoc Tukey’s test was used to evaluate differences among the five groups (control; MB/carrier pH 7.4; MB pH 7.4; MB/carrier pH 5.6; MB pH 5.6). The Statistics 8.0 software was used (P

Achyrocline flaccida aqueous extract was obtained by macerating wildflowers. The phytochemical profile present in the A. flaccida aqueous extract was elucidated by HPLC-ESI-MS/MS. Toxicity was evaluated in vitro by comet assay in peripheral blood mononuclear cells (PBMCs) and in vivo using Caenorhabditis elegans as a model. The antioxidant activity was also evaluated, and antimycobacterial activity was assessed by the broth microdilution method. The compounds present in the aqueous extract mainly belonged to the flavonoid class (89%). The concentrations that showed protective effects in C. elegans against oxidative stress and antimycobacterial activity had no toxic effects. The antimycobacterial activity test demonstrated that the concentration of 1,560 µg mL−1 inhibited the growth and eradication of the mycobacterial tested strains. Based on our findings, the A. flaccida aqueous extract presents a viable potential in developing new phytotherapeutic drugs against mycobacteria of clinical relevance.

The synthesis of 3-alkyl-5-trifluoromethyl-1H-pyrazole-1-carboxamides, 3-alkyl5-trifluoromethyl-1H-pyrazole-1-thiocarboxamides, and 2-(3-alkyl-5-trifluoromethyl-1H-pyrazol1-yl)-thiazoles derivatives are reported. [3 + 2] cyclocondensations for a series of long alkyl chain 1,1,1-trifluoro-4-methoxyalk-3-en-2-ones and semicarbazide or thiosemicarbazide were carried out in ethanol, an eco-friendly medium. The series of trifluoromethyl-1H-pyrazol-1-ylthiocarboxamide following a [3 + 2] cyclocondensation with 2-bromoacetophenone were converted into two series of 2-(3-alkyl-5-trifluoromethyl-1H-pyrazol-1-yl)-thiazoles. Good yields (69-96%) of the isolated products were obtained. The structures of the new long alkyl chain 1H-pyrazoles and 2-(1H-pyrazol-1-yl)thiazoles were characterized using 1 H, 13C, and 19F nuclear magnetic resonance (NMR) spectroscopy and electrospray ionization tandem mass spectrometric (ESI MS/MS) data. Moreover, some of the products were evaluated for their antimicrobial activity against Gram-negative Escherichia coli American Type Culture Collection (ATCC) 35218, Salmonella enteritidis ATCC 13076, and Pseudomonas aeruginosa ATCC 15692; and Gram-positive Staphylococcus aureus ATCC 6538, methicillin resistant Staphylococcus aureus (MRSA clinical isolate), Streptococcus sp. (clinical isolate), and Candida albicans ATCC 14053 and Candida krusei ATCC 6258 fungi. All the tested 1H-pyrazoles exhibited antibacterial and antifungal activities at the tested concentrations. The compounds from series 4 were found to be powerful against MRSA.

This study investigated the chemical constituents of

Natural products are still a promising source of bioactive molecules. Food and Drug Administration data showed that approximately 49% of the approved molecules originate naturally or chemically-resemble these substances, of which more than 70% are being used in anticancer therapy. It is noteworthy that at present there are no scientific studies to prove the effectiveness and safety of a number of plants used in folk medicine such as in the case of

In this work, we report the antimicrobial photoinactivation using tetra-cationic porphyrins with peripheral palladium(II) complexes against bacterial strains. Two different isomeric Pd(II)-porphyrins were used and applied in PDI experiments in the strains Gram negative bacilli Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa and a Gram positive cocci Staphylococcus aureus. All experiments are conducted with adequate concentration (without aggregation) of photosensitizer under white-light irradiation over 2 h . We showed that the most effective PS caused a significative reduction in the concentration of viable bacteria. The present results show that porphyrin with positively charge at meta position (3-PtTPyP) are more efficient photosensitizer against bacteria strains. In this way, tetra-cationic porphyrins might be promising antibacterial PDI agents with potential applications in clinical cases.

Naringin is a flavonoid widely known for its pharmacological properties, such as: anti-inflammatory and antioxidant ones, being an ally to avoid oxidative damage. Although naringin is an active easily found in citrus fruits, it has low bioavailability, biodistribution and also undergoes biotransformation in naringenin, limiting the described effects. The use of nanocapsules as drug carriers may increase solubility, improve biodistribution, impede the biotransformation thereof, and thus could improve the performance of naringin for use in treating neurological diseases. Therefore, the objective of this work is to produce a nanocapsule containing naringin, validate an analytical method by RP-HPLC to determination of the drug in nanoparticle and evaluate the toxicity. To that end, the blank nanocapsules (NB, without the drug) or naringin-loaded nanocapsules (NN) at the concentration of 2 mg/mL were prepared by interfacial deposition of the preformed polymer and the quantification of naringin by HPLC. Toxicity of the formulations was evaluated in vitro in rat hippocampal slices and in vivo models with C. elegans and Danio rerio (zebrafish). The analytical parameters evaluated (linearity, limit of detection and quantification, specificity, precision, accuracy and robustness) indicated adequate method to assay of naringin in nanocapsules by HPLC. There was no indication of toxicity by the nanocapsules in the evaluated biological assays.

A simple, rapid, low-cost method was proposed for the imaging of Pseudomonas aeruginosa biofilms on metallic surfaces using an infrared camera. Stainless steel coupons were cooled to generate a thermal gradient in relation to biofilm for active thermography (AT). Both cooling and image acquisition times were optimized and the images obtained with AT were compared with those from scanning electron microscopy. A free software (Thermofilm) was developed for image processing and the results were compared with the software ImageJ, with good agreement (from 87.7 to 103.8%). Images of coupons treated with sanitizer (peracetic acid) were obtained to show the applicability of the proposed method for biofilm studies. All analytical steps could be performed in 3 min in a noncontact, nondestructive, low-cost, portable, and easy-to-use way.