The aim of this study was to evaluate the antioxidant, antibacterial, and antibiofilm activities of nerolidol. The antioxidant activity of nerolidol was determined using the total antioxidant ...activity method. Antibacterial activity was performed using the microdilution method to determine the minimum inhibitory concentration (MIC) against seven standard strains of the ATCC and four bacterial clinical isolates with a resistance profile, following the Clinical and Laboratory Standards Institute (CLSI). The antibiofilm activity of nerolidol was performed using the crystal violet method. The results of the antioxidant test revealed a total antioxidant activity of 93.94%. Nerolidol inhibited the growth of
Staphylococcus aureus
(MIC = 1 mg/mL),
Streptococcus mutans
(MIC = 4 mg/mL),
Pseudomonas aeruginosa
(MIC = 0.5 mg/mL), and
Klebsiella pneumoniae
(MIC = 0.5 mg/mL). For clinical isolates, nerolidol showed an inhibitory potential against multidrug-resistant
P. aeruginosa
,
K. pneumoniae
carbapenemase (MIC = 0.5 mg/mL), methicillin-susceptible
S. aureus
(MIC = 2 mg/mL), and methicillin-resistant
S. aureus
(MIC = 2 mg/mL). Nerolidol showed similar antibacterial activity against ATCC strains and hospital clinical isolates with resistance profile, suggesting that even though these strains are resistant to antibiotics, they are still sensitive to nerolidol. Nerolidol exerted a dose-dependent effect on the inhibition of biofilm formation, even at subinhibitory concentrations. Nerolidol inhibited bacterial biofilms of ATCC strains at a rate ranging from 51 to 98%, at concentrations ranging from 0.5 to 4 mg/mL. For clinical bacterial isolates, biofilm inhibition ranged from 6 to 60%. Therefore, the present study showed the antioxidant, antibacterial, and antibiofilm properties of nerolidol.
Pseudobombax marginatum, popularly known as "embiratanha," is widely used by traditional communities as anti-inflammatory and analgesic agent. This study aimed to determine the phytochemical profile ...as well as cytotoxicity, acute oral toxicity, genotoxicity, and mutagenicity attributed to exposure to aqueous (AqEx) and ethanolic (EtEx) extracts of embiratanha bark. Phytochemical screening was conducted using thin-layer chromatography (TLC). Cell viability was analyzed using MTT assay with human mammary gland adenocarcinoma (MDA-MB-231) and macrophage (J774A.1) cell lines, exposed to concentrations of 12.5, 25, 50, or 100 µg/ml of either extract. For acute oral toxicity, comet assay and micronucleus (MN) tests, a single dose of 2,000 mg/kg of either extract was administered orally to Wistar rats. TLC analysis identified classes of metabolites in the extracts, including cinnamic acid derivatives, flavonoids, hydrolyzable tannins, condensed tannins, coumarins, and terpenes/steroids. In the cytotoxicity assay, the varying concentrations of extracts derived from embiratanha induced no significant alterations in the viability of MDA-MB-231 cells. The lowest concentration of EtEx significantly increased macrophage J774A.1 viability. However, the higher concentrations of AqEx markedly lowered macrophage J774A.1 viability. Animals exhibited no toxicity in the parameters analyzed in acute oral toxicity, comet assay, and MN tests. Further, EtEx promoted a significant reduction in DNA damage index and DNA damage frequency utilizing the comet assay, while the group treated with AqEx exhibited no marked differences. Thus, data demonstrated that AqEx or EtEx of embiratanha may be considered safe at a dose of 2,000 mg/kg orgally under our experimental conditions tested.
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•A novel lectin was successfully purified from Dypsis decaryi seeds.•A glycoprotein was purified from D. decaryi by ion-exchange and gel filtration chromatography.•The activity of ...DdeLL was not inhibited by all monosaccharides tested.•DdeLL is stable within a pH range of 5.0–7.5 and temperature upto 100 °C.•DdeLL exhibits high acaricide activity against Rhipicephalus (Boophilus) microplus.
A novel thermostable lectin, called DdeL, was obtained from the extract of Dypsis decaryi seeds. The extract of the tegument had the highest specific hemagglutination activity (196.03 HA/mg) when subjected to 70% acetone precipitation and purification on Superdex G-75 and exhibited a single peak (38.8 kDa). DdeL, a glycoprotein, had agglutinated blood types and was inhibited only by glycoproteins (albumin, azoalbumin and azocasein). Biochemical properties showed higher activity at acid pH (pH 5–6) than alkaline pH and activity was observed at all temperatures (up to 100 °C). DdeL exhibited bacteriostatic activity against Providencia stuartii and Staphylococcus aureus ATCC 29213, which presented minimum inhibitory concentration (MIC) of 750.0 μg/mL. When tested against fungi Sporotrichum schenckii and Candida albicans showed excellent MIC50 at 0.52 and 2.06 μg/mL, respectively. DdeL showed no cytotoxic effect on macrophages (J774.A1) and renal embryonic cells (HEK-293). in vitro acaricide activity of DdeL against Rhipicephalus showed high activity in low concentration. Therefore, we conclude that DdeL may be of interest for potential biotechnological applications.
This study aimed to characterize the phytochemical profile of bark and leaves aqueous extract Commiphora leptophloeos, and conduct in vivo and in vitro assays to determine the presence of any ...toxicological consequences due to exposure. The phytochemical analysis was carried out using high-performance liquid chromatography (HPLC). The antioxidant activity was estimated utilizing DPPH free radical scavenging and phosphomolybdenum assays. Cell viability was measured by the MTT method on J774 and human adenocarcinoma cells, which were treated with concentrations of 12,5, 25, 50, 100 or 200 µg/ml of both extracts. Acute oral toxicity, genotoxicity, and mutagenicity assays were determined using a single oral dose of 2000 g/kg in male Swiss albino mice (Mus musculus). Biochemical analysis of the blood and histological analyses of the kidneys, liver, spleen, pylorus, duodenum and jejunum were undertaken. Genotoxicity and mutagenicity were determined utilizing blood samples. Gallic acid, catechin, and epicatechin were identified in the bark and chlorogenic acid in leaves. Data demonstrated a high content of phenolic compounds and flavonoids associated with significant antioxidant potential. No significant signs in damage or symptoms of toxicity were detected. No marked reduction in cell viability was found at lower concentrations tested. On histomorphometry, only the gastrointestinal organs exhibited significant difference. Renal hepatic and blood parameters were within the normal range. No apparent signs of toxicity, genotoxicity, mutagenicity or cytotoxicity were found in vivo and in vitro experiments.
The objective of this study was to determine the cytotoxicity of organic extracts of P. moniliformis in vitro and identify the acute toxicity and genotoxicity in vivo. The leaves were extracted using ...three organic solvents (cyclohexane EP1, ethyl acetate EP2, and methanol EP3). Phytochemical qualitative analysis was performed by thin layer chromatography (TLC). Cytotoxicity tests were performed on human embryonic kidney (HEK) cells and J774 murine macrophages. Acute toxicity in mice was measured after intraperitoneal (ip) administration of 2000 mg/kg, while evaluation of genotoxicity and mutagenicity were assessed using the comet assay and the micronucleus (MN) test, respectively. The TLC analysis of the extracts revealed the presence of flavonoids, triterpenes, steroids, and saponins. In the cytotoxicity assay, extracts EP1 and EP3 altered proliferation of HEK cells, and all organic extracts increased the viability of J774 cells. In the toxicity tests, no deaths or behavioral alterations were observed in mice exposed to the acute dose of the extracts. Although some extracts led to changes in hematological and histological parameters, these results did not indicate physiological changes. In relation to the MN test and comet assay, no significant changes were detected in the DNA of the animals tested with the extracts EP1, EP2, and EP3. Thus, extracts of P. moniliformis were not considered to be toxic and did not induce formation of MN or damage to cellular DNA in the genotoxicity tests.
Despite numerous scientific advances, cancer continues to be one of the main causes of death in the world. This situation has driven the search for promising molecules. Lichen substances have been ...widely described for their pharmacological potential.
The present study evaluated the antitumour potential of a depsidone isolated from
- salazinic acid (SAL) - through
,
and
studies.
The molecule was isolated from the acetonic extract of the lichen and recrystallized in acetone. The macrophage J774, sarcoma-180 and MDA-MB-231 cell lines were used for the MTT cytotoxicity assay. The antitumor assay used a murine model (Swiss albino mice) with sarcoma-180. The animals were treated for seven consecutive days with doses of SAL (25 and 50 mg/kg) and 5-fluorouracil (20 mg/kg).
Its purity was determined using high-performance liquid chromatography (94%), and its structure was confirmed by H
and C
nuclear magnetic resonance. SAL was not considered toxic to cancer cell lines, showing cell viability rates of 79.49 ± 4.15% and 86.88 ± 1.02% for sarcoma-180 and MDA-MB-231, respectively. The tumour inhibition rate was greater than 80% in the animals treated with SAL and 65% for those that received 5-fluorouracil. Simulations of molecular dynamics to estimate the flexibility of the interactions between human thymidylate synthase and derivatives of SAL and 5-fluorouracil revealed that SAL exhibited greater enzymatic interaction capacity, with highly favourable energy, compared to 5-fluorouracil.
The present results demonstrate the potential of salazinic acid as a tumour inhibition agent.
Thrombosis is characterized by the pathological formation of fibrin clots within a blood vessel, leading to the obstruction of blood flow. Fibrinolytic enzymes from microorganisms have been shown to ...be more efficient and safer in dissolving clots. Then, this study aimed to evaluate the cell growth and fibrinolytic enzyme production of Tetradesmus obliquus under different cultivation conditions. T. obliquus grew under autotrophic and mixotrophic conditions using different concentrations of corn steep liquor (0.25 ≤ CSL ≤ 4.00%). The cells were concentrated and lysed via two different methods (sonication or homogenization) to trigger the release of the enzyme. It was precipitated via acetone or ammonium sulfate additions and purified using ion exchange chromatography. The highest biomass productivity (Px = 130 ± 12.8 mg∙L−1day−1), specific growth rate (µmax = 0.17 ± 0.00 day−1), and fibrinolytic activity (391 ± 40.0 U∙mg−1) was achieved on a mixotrophic cultivation at a 0.25% CSL concentration. The results showed that the homogenizing method had better performance in the release of enzyme, and the precipitation with acetone obtained the highest fibrinolytic activity (567 ± 49.3 U∙mg−1). The purified enzyme showed a specific activity of 1221 ± 31 U∙mg−1 and a molecular mass of 97 kDa. So, the fibrinolytic enzyme from T. obliquus had higher activity when compared to the other fibrinolytic enzymes, being a potential source for the development of therapeutic agents in thrombosis treatment. Additional studies are needed to investigate the biochemical properties and biological profile of this enzyme.
This work reports the immobilization of a fibrinolytic protease (FP) from Mucor subtilissimus UCP 1262 on Fe3O4 magnetic nanoparticles (MNPs) produced by precipitation of FeCl3·6H2O and FeCl2·4H2O, ...coated with polyaniline and activated with glutaraldehyde. The FP was obtained by solid state fermentation, precipitated with 40–60% ammonium sulfate, and purified by DEAE-Sephadex A50 ion exchange chromatography. The FP immobilization procedure allowed for an enzyme retention of 52.13%. The fibrinolytic protease immobilized on magnetic nanoparticles (MNPs/FP) maintained more than 60% of activity at a temperature of 40 to 60 °C and at pH 7 to 10, when compared to the non-immobilized enzyme. MNPs and MNPs/FP did not show any cytotoxicity against HEK-293 and J774A.1 cells. MNPs/FP was not hemolytic and reduced the hemolysis induced by MNPs from 2.07% to 1.37%. Thrombus degradation by MNPs/FP demonstrated that the immobilization process guaranteed the thrombolytic activity of the enzyme. MNPs/FP showed a total degradation of the γ chain of human fibrinogen within 90 min. These results suggest that MNPs/FP may be used as an alternative strategy to treat cardiovascular diseases with a targeted release through an external magnetic field.
•Immobilization is efficient to fibrinolytic enzyme from Mucor subtilissimus.•The immobilization process ensured the coupling of 52.13% of the protease.•The immobilized enzyme maintained more than 60% of activity compared to the free.•This immobilized enzyme an alternative for the treatment of cardiovascular diseases.
Fibrinolytic enzymes from microorganisms have been investigated by their potential application as thrombolytic agents. Previous studies show that the fibrinolytic enzyme from Arthrospira platensis ...(FEAP) is stable at human physiological temperature (<50 °C) and pH (6.0–10.0). Understanding of the kinetic/thermodynamic characteristics is important to make the advances for industrial applications feasible. Therefore, a kinetic/thermodynamic analysis of FEAP was performed on the fibrin hydrolysis and enzyme thermoinactivation. Results showed two Michaelis-Menten-type profiles with two plateaus, revealing high affinity for the substrate with low Michaelis constant values (0.02 and 6.37 µg∙mL−1). The activation energy of the hydrolysis catalyzed by FEAP and the standard enthalpy variations of reversible enzyme unfolding were 49.65 and 107.01 kJ∙mol−1, respectively. When the temperature increased from 40 to 70 °C, the deactivation rate constant increased from 0.0050 to 0.0134 min−1, while the half-life decreased from 138.62 to 51.72 min. These results allowed to estimate the activation energy (E*d = 27.50 kJ∙mol−1), enthalpy (24.64 ≤ ΔH*d ≤ 24.89 kJ∙mol−1), entropy (−209.86 ≤ ΔS*d ≤ −210.10 J∙mol−1∙K−1), and Gibbs free energy (90.61 ≤ ΔG*d ≤ 96.74 kJ∙mol−1) of thermal denaturation. The data suggest that FEAP is a promising and thermostable biocatalyst that could be exploited for industrial applications.
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•Kinetic/thermodynamic parameters were defined for the fibrinolytic enzyme from Arthrospira platensis.•Fibrinolytic enzyme showed two different Michaelis-Menten kinetic profiles.•Substrate binding and catalytic efficiency were improved at lower fibrin concentration (<7.81 µg/mL).•The enzyme is more stable at lower temperatures.
Syagrus coronata, popularly known as licuri, is a palm native to caatingas. The fixed oil extract of licuri nuts is used by the population of Northeast Brazil for therapeutic purposes, including as ...an antifungal, anti-inflammatory, and a cicatrizant agent. However, there is no scientific information on the possible harmful health effects of the oil and hence its medicinal usability is unknown.
We aimed to analyze the biological safety and possible antioxidant activity of fixed S. Coronata oil.
Chemical analysis of the oil was performed using gas chromatography with flame ionization detection (CG-FID). The cytotoxicity of varying concentrations of the oil (12.5, 25, 50, 100, and 200 μg/mL) was evaluated using the tetrazolium reduction assay in three cell lines: HEK-293 kidney embryonic cells, J774.A1 macrophages, and the tumor line Sarcoma-180 (S-180). Oral toxicity, genotoxicity, and mutagenicity tests were performed in mice which were administered a single dose of 2000 mg/kg of fixed licuri oil, by gavage. For acute toxicity tests, changes in blood and biochemical parameters, behavior, and weight were analyzed; histomorphometric analyses of the liver, kidney, and spleen were also performed. The comet assay and micronucleus (MN) test were performed to analyze genotoxicity. The antioxidant potential was assessed by the total antioxidant capacity (AAT) and DPPH elimination activity.
Licuri oil consists predominantly of saturated fatty acids, and lauric acid is the major compound. The highest concentrations of the oil showed low levels of cytotoxicity; however, LC50 was not reached in any of the tests. The acute toxicity study did not reveal any evidence of adverse effects in animals treated with oil; biochemical investigation of blood showed a decrease in blood concentration of total proteins and uric acid. The kidneys, spleen, and liver showed no morphological changes indicative of a pathological process. Genotoxic or mutagenic activity was not detected through both the comet assay and MN test. In addition, the oil showed low antioxidant activity in both methods.
Licuri oil from the stem of S. coronata did not present significant toxic effects as well as absence of genetic damage when administered orally. Future studies are needed to investigate its pharmacological potential.
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•In Brazil, fixed oil from the nuts of S. coronata is used for therapeutic purposes.•Its safety for human consumption needs to be established.•We performed in vitro cytotoxicity, in vivo acute toxicology, and genotoxicity assays.•The oil did not induce cell death, nor morphological alterations in analyzed tissues.•The oil has no mutagenic nor genotoxic effects.
