RESUMO
Fungi are an important source of polysaccharides (PSH) and phenolic compounds (PC). Numerous studies have highlighted the beneficial effects of fungal consumption, but the impact of submerged cultivated mycelia (M) and filtrate (F) has not been fully investigated. We aimed to investigate the cytotoxic activity of isolated crude PSH and exopolysaccharides (ePSH) of submerged cultivated M and F of edible Coprinus comatus and Coprinellus truncorum species. Both PSH and ePSH exhibited significant cytotoxic activity towards HepG2 cancer cells of human origin (three-way ANOVA). The C. truncorum PSH/ePSH was more efficient inducing maximal reduction in cell viability (≈50% at 450 µg/mL) after 24 h while C. comatus PSH/ePSH needed 72 h to reach similar effect (≈60% at 450 µg/mL). Partial least square regression (PLSR) analysis indicated that specific phenolic composition of the PSH/ePSH could be responsible for the difference in their activity.
Assuntos
Polissacarídeos , Polissacarídeos/química , Polissacarídeos/farmacologia , Espectroscopia de Infravermelho com Transformada de Fourier , Cromatografia Gasosa-Espectrometria de Massas , Linhagem Celular Tumoral , Humanos , Sobrevivência Celular/efeitos dos fármacosRESUMO
A new protein detector of 2,4,6-trinitrophenol (TNP, picric acid) is concisely reported herein for the first time. In brief, the gene of a specific scFv fragment, namely 3.5, was fused separately with the gene of ß-lactamase and, subsequently, expressed in the periplasmic space of Escherichia coli, affording in such a way the recombinant fusion protein 3.5-scFv-ß-lactamase. This bifunctional immunoreagent tool was further convincingly demonstrated for being capable to directly detect trinitrophenol-tris(hydroxymethyl)aminomethane (TNP-Tris), a less toxic TNP derivative of choice compared to TNP itself, with competitive sensitivity (50 ± 2 fmol or 175 ± 6 pg/mL).[Formula: see text].
Assuntos
Substâncias Explosivas , beta-Lactamases , Região Variável de Imunoglobulina/genética , Região Variável de Imunoglobulina/metabolismo , Nitratos , Picratos , beta-Lactamases/genética , beta-Lactamases/metabolismoRESUMO
This study aimed to develop a fast and reliable protocol for Trinitrophenol-Tris(hydroxymethyl)aminomethane (TNP-Tris) detection applying a ß-lactamase-fusion protein of choice, the natural product-based immunoreagent tool of competitive sensitivity developed herein for the first time. Since the fusion protein 11B3-scFv-ß-lactamase is constructed from a scFv-antibody (11B3) linked to an enzyme (ß-lactamase), the step related to the use of secondary antibody in the enzyme-linked immunosorbent assay (ELISA) is completely omitted. Indeed, this fusion protein itself serves both as binding mean of the antigen model and detecting agent, due to the presence of the naturally occurring enzyme. In such a way, it actually affords the one-step TNP-Tris detection reaching a promising LOD value of 45 ± 2 fmol or 157 ± 6 pg/mL. Taken all together, the current protocol does represent much cheaper and significantly less-time consuming alternative compared both to the recombinant antibodies and recombinant phages, previously designed means in our labs for the same purpose.
Assuntos
Produtos Biológicos , Ensaio de Imunoadsorção EnzimáticaRESUMO
The cytotoxicity of the methanol extract of the freshwater sponge Ochridaspongia rotunda (Arndt, 1937) (Malawispongiidae) was evaluated by MTT assay at in vitro conditions against three brain tumour cell lines (Neuro-2A, U-251 MG and U-87 MG). The extract was actually found to be most effective against the malignant glioma U-251 MG cells reaching a promising IC50 value of 1.87 ± 0.09 µg/mL at 96 h. However, it exhibited only a bit of cytotoxicity (IC50 321.14 ± 11.29 µg/mL, 96 h) towards the normal cells. Also, this sponge extract was 5-fold more selective for U-251 MG versus U-87 MG cells. Finally, monitoring genotoxicity at chromosomal level using the micronucleus test practically revealed lack of any significant toxicity of O. rotunda extract, compared to doxorubicin.
Assuntos
Produtos Biológicos/uso terapêutico , Glioblastoma/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Ácidos Graxos Insaturados/farmacologia , Glioblastoma/patologia , Humanos , Metanol/química , Testes para Micronúcleos , Extratos Vegetais/farmacologia , Poríferos/química , Espectroscopia de Prótons por Ressonância MagnéticaRESUMO
This study was focused on in vitro cytotoxicity screening of the raspberry seeds methanol extract towards a number of cancer cell lines of human origin. The tested extract at the preferred concentrations (IC50 <30 µg/mL) inhibited only the growth of the lung cancer A-549 cells (IC50 = 14.07 ± 0.96 µg/mL). At the same time, it was practically inactive (IC50 >300 µg/mL) and non-mutagenic towards normal MRC-5 lung cells. Finally, the extract potently scavenged both OH· (IC50 = 20.11 ± 1.77 µg/mL) and O2-· (IC50 = 47.23 ± 3.82 µg/mL), the free radicals of proved relevance for cancer pathophysiology. Though seeds were enriched with phenolic compounds (TPC = 5.21 ± 0.07 mg GAE/g), anthocyanins were present in traces only (TAC = 0.07 ± 0.003 mg cyn-3-glu/g), while flavonoids were not detected at all. This is the first report on anti-lung cancer potential of the seeds of any soft fruit.
Assuntos
Neoplasias Pulmonares/patologia , Extratos Vegetais/farmacologia , Rubus/química , Sementes/química , Antioxidantes/farmacologia , Linhagem Celular Tumoral , Humanos , Concentração Inibidora 50RESUMO
This study focuses on the impact of low and high temperature regimes on the chemical composition and antioxidativity of low sugar berry fruits jellies. High quality fruits (strawberry, raspberry and blackberry) were collected from Western Serbia region, quite well recognised both nationally and internationally due to an extremely well developed practice in growing berry fruits. The obtained results have clearly indicated the importance of low temperature regime for enriched contents of both total phenolics and anthocyanins followed by an enhanced antioxidativity. Rubus fruticosus L. Cacak Thornless cultivar, the only autochthonous berry fruit variety screened herein, may be firmly recommended as a raw material for industrial production of low sugar blackberry jellies with exceptional characteristics. This innovative procedure of preparing berry fruit jellies have encompassed the application of low temperature regime (55 °C), lower content of sugar (40%), seeds separation from jellies followed by no use of pectin throughout the whole process.
Assuntos
Indústria de Processamento de Alimentos/métodos , Fragaria/química , Frutas/química , Rubus/química , Antocianinas/análise , Antocianinas/química , Antioxidantes/análise , Géis , Fenóis/análise , Fenóis/química , Sérvia , Açúcares/análise , TemperaturaRESUMO
This study was focused on in vitro screening of the total antioxidant activity of the selected extracts of the mushroom Coprinus comatus and quinic acid, one of their antidiabetic ingredients, by an uncommon electrochemical assay. Indeed, direct current (DC) polarographic HydroxoPerhydroxo Mercury(II) Complex (HPMC) assay based on decrease of anodic limiting current originating from HPMC formation in alkaline solutions of hydrogen peroxide at potential of mercury dissolution, observed upon gradual addition of antioxidants, was applied herein for the estimation of the natural products' antioxidativity. Quinic acid was found to exhibit most promising antioxidant potential (4.0 ± 0.2%µL-1) being ≈ 2-fold more active than the screened C. comatus extract samples. Actually, such a finding puts some light on the antioxidativity of cyclic polyols, well understimated class of organic compounds, compared to aromatic (poly)phenolics. As a low cost, easy-to handle and accurate this polarographic assay may be thoroughly recommended for much broader use. [Formula: see text].
Assuntos
Antioxidantes/análise , Coprinus/química , Hipoglicemiantes/análise , Polarografia , Ácido Quínico/análise , Antioxidantes/química , Produtos Biológicos/análise , Hipoglicemiantes/química , Micélio/química , Ácido Quínico/químicaRESUMO
Studies on the identification, properties and function of chitin in sponges (Porifera), which are recognized as the first multicellular organisms on Earth, continue to be of fundamental scientific interest. The occurrence of chitin has so far been reported in 21 marine sponge species and only in two inhabiting fresh water. In this study, we present the discovery of α-chitin in the endemic demosponge Ochridaspongia rotunda, found in Lake Ohrid, which dates from the Tertiary. The presence of chitin in this species was confirmed using special staining, a chitinase test, FTIR, Raman and NEXAFS spectroscopy, and electrospray ionization mass spectrometry (ESI-MS). In contrast to the case of marine sponges, chitin in O. rotunda has been found only within its holdfast, suggesting a role of chitin in the attachment of the sponge to the hard substratum. Isolated fibrous matter strongly resemble the shape and size of the sponge holdfast with membrane-like structure.
Assuntos
Quitina/química , Quitina/metabolismo , Poríferos/química , Poríferos/metabolismo , AnimaisRESUMO
Autochthonous variety 'Prokupac' (Vitis vinifera L.) red wines with the addition of selected aromatic herbs (Artemisia absinthium, Cinnamomum zeylanicum, Glycyrrhiza glabra and Pimpinella anisum L.) were evaluated for chemical composition and biological activity. Furthermore, their polyphenolic profiles were determined by HPLC-DAD. While total phenolic (TPC) and flavonoid (TFC) contents, along with antioxidant activity, were significantly higher in all examined wine samples, the content of total anthocyanins was decreased, compared to a pure 'Prokupac' wine, used as the control one. On the other hand, 'Prokupac' wines with the addition of cinnamon (S. zeylanicum) and wormwood (A. absinthium) displayed highest antioxidant (EC50, 0.021 ± 0.001 mg/mL) and antibacterial (Enterococcus faecalis, MIC/MBC 15.63/62.50 µL/mL) activities, respectively. The study showed that 'Prokupac' wine with the addition of cinnamon was significantly enriched both with TPC and TFC, compared to control wine (p < 0.05), which may contribute to its market potential.
Assuntos
Antibacterianos/análise , Antioxidantes/análise , Vitis/química , Vinho/análise , Antocianinas/análise , Antibacterianos/isolamento & purificação , Antioxidantes/isolamento & purificação , Artemisia , Cinnamomum zeylanicum , Flavonoides/análise , Fenóis/análise , Fenóis/químicaRESUMO
In vitro acetylholinesterase (AChE) inhibitory activity of an autochthonous sample of the mushroom Coprinus comatus (encompassing fruiting body FB, mycelia M and filtrate F from the submerged cultivation) was the subject of this study. C. comatus F extract exhibited rather potent anti-AChE activity (73.0 ± 1.5%) at in liquid conditions, comparable to those of the conventional drug donepezil (80.6 ± 1.4%). Also, the same extract exhibited high anti-AChE activity (1 µg) in solid. While its FTIR spectrum indicated the presence of phenolic compounds, quercetin (28.1 µg g-1 d.w.) was found to affect the observed bioactivity (59.8 ± 0.9%). This is the first report of profound anti-AChE activity of any C. comatus extract, a medicinal mushroom that has been successfully cultivated in P.R. China, due to the demanding needs of food industry.
Assuntos
Coprinus/química , Fármacos Neuroprotetores/isolamento & purificação , Esterol Esterase/antagonistas & inibidores , Agaricales , China , Fenóis/análise , Quercetina/isolamento & purificação , Quercetina/farmacologiaRESUMO
The liposomal integration method, in conjunction with electron paramagnetic resonance (EPR) spectroscopy, has been presented for the investigation of antioxidant activity of selected water-insoluble compound towards biologically relevant free radicals. This method was applied to avarol, a sesquiterpenoid hydroquinone isolated from the marine sponge Dysidea avara. The antioxidant activity of water-insoluble avarol towards â¢OH, O2â¢- and NO⢠radicals was attained by its incorporation into the DPPC liposomes bilayer, and towards ascorbyl radicals in the organic solvent. Avarol's activity towards â¢OH, O2â¢-, NO⢠and ascorbyl radicals was 86.2%, 50.9%, 23.6% and 61.8%, respectively, showing its significant radical scavenging potential.
Assuntos
Antioxidantes/farmacologia , Radicais Livres/antagonistas & inibidores , Sesquiterpenos/farmacologia , Animais , Antioxidantes/química , Antioxidantes/isolamento & purificação , Dysidea/química , Lipossomos , Estrutura Molecular , Sesquiterpenos/química , Sesquiterpenos/isolamento & purificação , Solubilidade , Água/químicaRESUMO
Compared to plants, nowadays mushrooms attract more attention as functional foods, due to a number of advantages in manipulating them. This study aimed to screen the chemical composition (fatty acids and phenolics) and antioxidant potential (OHâ¢, 2,2-diphenyl-1-picrylhydrazyl (DPPHâ¢) and ferric reducing ability of plasma (FRAP)) of two edible mushrooms, Coprinus comatus and Coprinellus truncorum, collected from nature and submerged cultivation. Partial least square regression analysis has pointed out the importance of some fatty acids-more precisely, unsaturated fatty acids (UFAs) followed by fatty acids possessing both short (C6:0 and C8:0) and long (C23:0 and C24:0) saturated chains-and phenolic compounds (such as protocatechuic acid, daidzein, p-hydroxybenzoic acid, genistein and vanillic acid) for promising anti-OHâ¢, FRAP and anti-DPPH⢠activities, respectively. However, other fatty acids (C16:0, C18:0 and C18:3n3) along with the flavonol isorhamnetin are actually suspected to negatively affect (by acting pro-oxidative) the aforementioned parameters, respectively. Taken together, design of new food supplements targeting oxidative stress might be predominantly based on the various UFAs combinations (C18:2n6, C20:1, C20:2, C20:4n6, C22:2, C22:1n9, etc.), particularly if OH⢠is suspected to play an important role.
RESUMO
The elemental profile of the leaf samples of three Cabernet Franc clone candidates recently developed in Serbia (Nos. 02, 010 and 012) obtained in the last phase of clonal selection was examined within this study by ICP-OES. Optimal content of Al, Fe, Cu and Zn, the metals well known for their links with a number of neurodegenerative disorders including Alzheimer's disease, has actually highlighted the potential of the leaf sample No. 10 to afford a novel food supplement of natural origin possessing well balanced metal ingredients. Furthermore, lower content of some other elements (Cr, Mn, Ni and Pb) thoroughly supports such a claiming. Finally, the most favourable K/Na ratio observed for the aforementioned sample points out its likely cardioprotectivity. However, two other Cabernet Franc clone candidates might also be recommended for breeding in the same or similar viticultural conditions, since all obtained values were below toxic ones for human consumption.
Assuntos
Metais/análise , Folhas de Planta/química , Vinho/análise , Células Clonais , Análise de Perigos e Pontos Críticos de Controle , Humanos , SérviaRESUMO
Nowadays, the mangrove biome is considered to be a profound resource of natural products usually possessing cytotoxicity of a broader range. Covering the period 2007-2015, a total of 21 new naturally occurring compounds has stood out. For example, xylogranin B and swietephragmin C were found to exhibit very potent cytotoxic activity against the colon HCT-116 cells reaching IC50 values of 0.05 and 0.06 µM, respectively. Bearing in mind the efficacy of the majority compounds in the preliminary in vitro screens, these studies should be expanded to both ex vivo and in vivo screens including the evaluation of the relevant toxicological profiles.
Assuntos
Produtos Biológicos/isolamento & purificação , Produtos Biológicos/farmacologia , Áreas Alagadas , Antineoplásicos Fitogênicos/farmacologia , Catecóis/isolamento & purificação , Catecóis/farmacologia , Células HCT116 , Humanos , Lignanas/isolamento & purificação , Lignanas/farmacologia , Limoninas/farmacologia , Micotoxinas/isolamento & purificação , Micotoxinas/farmacologia , Xantonas/isolamento & purificação , Xantonas/farmacologiaRESUMO
The polysaccharide (PSH) extracts from the edible mushroom species Coprinus comatus and Coprinellus truncorum were screened in liquid for their acetylcholinesterase inhibitory (AChE) activity. Both extracts were found to display inhibition of the aforementioned enzyme reaching similar IC50 values of 0.62 ± 0.07 and 0.61 ± 0.03 mg/mL, respectively. According to the means of FTIR spectroscopy, these PSH extracts mostly contained ß-glucans. However, the presence of some proteins and polyphenolics as minor ingredients were also detected. Compared with existing literature data for anti-AChE activity of the sugar samples, the findings within this study may be treated as a profound bioactivity. Consequently, this study puts some light on the possible use of the screened macrofungi in the palliative treatment of Alzheimer's disease.
Assuntos
Inibidores da Colinesterase/farmacologia , Coprinus/química , Polissacarídeos/farmacologia , Acetilcolinesterase/metabolismo , Agaricales/química , Inibidores da Colinesterase/isolamento & purificação , Proteínas Fúngicas/isolamento & purificação , Fenóis/isolamento & purificação , Polissacarídeos/isolamento & purificação , beta-Glucanas/isolamento & purificaçãoRESUMO
This study outlines antioxidant and anti-AChE activities of the polysaccharide (PSH) extract from the mushroom species Amanita strobiliformis. Both the presence of α and ß glucans within the aforementioned extract was recorded. PSH extract displayed a profound scavenging activity of OH radicals (IC50 value, 11.86 ± 0.59 µg/mL) and high potential for reduction of Fe3+ ions (174.11 ± 8.70 mg eq. AA/g d.w.) being almost 48- and 5-fold more effective than mannitol and butylated hydroxytoluene used as a positive control, respectively. Compared with galanthamine (0.001 µg), the same extract exhibited a moderate anti-AChE activity (10 µg) in solid. Since purified PSH extract exhibited higher bioactivity (IC50 value 7.27 ± 0.31 µg/mL, 197.68 ± 9.47 mg eq. AA/g d.w. and 0.1 µg, respectively), it can be predominantly ascribed to the polysaccharide compounds. A. strobiliformis PSH extract may be considered as a promising resource of potent bioactive polysaccharides of natural origin successfully addressing both oxidative stress and lack of acetylcholine.
Assuntos
Amanita/química , Antioxidantes/farmacologia , Inibidores da Colinesterase/farmacologia , Polissacarídeos Fúngicos/farmacologia , Hidroxitolueno Butilado/farmacologia , Avaliação Pré-Clínica de Medicamentos/métodos , Polissacarídeos Fúngicos/análise , Polissacarídeos Fúngicos/química , Radical Hidroxila/química , Radical Hidroxila/metabolismo , Manitol/farmacologia , Ácido Quínico/análise , SérviaRESUMO
Screens of antioxidant activity (AA) of various natural products have been a focus of the research community worldwide. This work aimed to differentiate selected samples of Merlot wines originated from Montenegro, with regard to phenolic profile and antioxidant capacity studied by survival rate, total sulfhydryl groups and activities of glutathione peroxidase (GPx), glutathione reductase and catalase in H2O2â»stressed Saccharomyces cerevisiae cells. In this study, DPPH assay was also performed. Higher total phenolic content leads to an enhanced AA under both conditions. The same trend was observed for catechin and gallic acid, the most abundant phenolics in the examined wine samples. Finally, the findings of an Artificial Neural Network (ANN) model were in a good agreement (r² = 0.978) with the experimental data. All tested samples exhibited a protective effect in H2O2â»stressed yeast cells. Pre-treatment with examined wines increased survival in H2O2â»stressed cells and shifted antioxidative defense towards GPxâ»mediated defense. Finally, sensitivity analysis of obtained ANN model highlights the complexity of the impact that variations in the concentrations of specific phenolic components have on the antioxidant defense system.
Assuntos
Antioxidantes/farmacologia , Polifenóis/farmacologia , Saccharomyces cerevisiae/citologia , Vinho/análise , Antocianinas/análise , Compostos de Bifenilo/química , Redes Neurais de Computação , Picratos/química , Análise de Componente Principal , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Compostos de Sulfidrila/metabolismoRESUMO
In vitro anti-tumour and anti-radical activities of the acetone extract of the freshwater sponge Ochridaspongia rotunda were the subject of this study. The extract was found to be highly cytotoxic to human lung tumour cell line A-549 reaching IC50 value of 5.01 ± 0.21 µg/mL. Indeed, it displayed only 2-fold less anti-tumour activity than doxorubicin (IC50 value 2.42 ± 0.13 µg/mL) used as a positive control. The same extract was also found to be almost 37-fold more selective against A-549 vs. MRC-5 (normal) lung cells, in difference to weak selectivity of doxorubicin (less than 3-fold). Its profound anti-DPPH radical activity comparable to that of quercetin (IC50 values 3.68 ± 0.19 and 3.14 ± 0.09 µg/mL, respectively) coupled with no signs of genotoxicity in the comet assay (MRC-5 cell line, vs. doxorubicin) has actually implicated the importance of this animal bioresource in searching for pharmaceutically useful bioactive compounds of natural origin.
Assuntos
Antineoplásicos/farmacologia , Poríferos/química , Extratos de Tecidos/farmacologia , Células A549 , Animais , Antineoplásicos/química , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Avaliação Pré-Clínica de Medicamentos/métodos , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/farmacologia , Água Doce , Humanos , Concentração Inibidora 50 , Extratos de Tecidos/químicaRESUMO
This review covers the 1994-2014 literature data published for anti-CNS tumour natural products isolated from marine sponges. The focus was on highly potent antitumourics. It describes only eight promising bioactives (screened on even less number of the respective cell lines) suggesting that brain tumours actually represent quite a hot topic for the natural product research community worldwide.
Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Neoplasias do Sistema Nervoso Central/tratamento farmacológico , Poríferos/química , Animais , Organismos Aquáticos , Produtos Biológicos/química , Produtos Biológicos/farmacologia , Neoplasias Encefálicas/tratamento farmacológico , Dioxanos/química , Dioxanos/farmacologia , Diterpenos/química , Diterpenos/farmacologia , Compostos Heterocíclicos de 4 ou mais Anéis/química , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Humanos , Pirimidinas/química , Pirimidinas/farmacologia , Terpenos/química , Terpenos/farmacologiaRESUMO
This study aimed to screen in vitro antitumour activity of the redox couple avarol/avarone against the human malignant glioma cell line U-251 MG for the first time. Compared both with avarol and positive controls used (temozolomide and doxorubicin), avarone was found to be the most active compound with IC50 value below 1 µM (IC50 0.68 ± 0.04 µM, 96 h). Considerable less DNA damage in the cells treated with avarol and avarone vs. doxorubicin (105 & 123% vs. 299%, respectively; untreated U-251 MG cells were used as a control, 100%), coupled with no sign of cytotoxicity against the normal human foetal lung fibroblast MRC-5 cells (IC50 > 100 µM), has actually pointed out the importance of this marine sesquiterpenoid quinone structure as a promising lead compound in development of novel brain chemotherapeutics.