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Breast cancer has a poor prognosis due to the toxic side effects associated with high doses of chemotherapy. Liposomal drug encapsulation has resulted in clinical success in enhancing chemotherapy tolerability. However, the formulation faces severe limitations with a lack of colloidal stability, reduced drug efficiency, and difficulties in storage conditions. Nanoarchaeosomes (NA) are a new generation of highly stable nanovesicles composed of the natural ether lipids extracted from archaea. In our study, we synthesized and characterized the NA, evaluated their colloidal stability, drug release potential, and anticancer efficacy. Transmission electron microscopy images have shown that the NA prepared from the hyperthermophilic archaeon Aeropyrum pernix K1 was in the size range of 61 ± 3 nm. The dynamic light scattering result has confirmed that the NA were stable at acidic pH (pH 4) and high temperature (70 °C). The NA exhibited excellent colloidal stability for 50 days with storage conditions at room temperature. The cell viability results have shown that the pure NA did not induce cytotoxicity in NIH 3T3 fibroblast cells and are biocompatible. Then NA were loaded with doxorubicin (NAD), and FTIR and UV-vis spectroscopy results have confirmed high drug loading efficiency of 97 ± 1% with sustained drug release for 48 h. The in vitro cytotoxicity studies in MCF-7 breast cancer cell lines showed that NAD induced cytotoxicity at less than 10 nM concentration. Fluorescence-activated cell sorting (FACS) results confirmed that NAD induced late apoptosis in nearly 92% of MCF-7 cells and necrosis in the remaining cells with cell cycle arrest at the G0/G1 phase. Our results confirmed that the NA could be a potential next-generation carrier with excellent stability, high drug loading efficiency, sustained drug release ability, and increased therapeutic efficacy, thus reducing the side effects of conventional drugs.
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The global impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its companion disease, COVID-19, has reminded us of the importance of basic coronaviral research. In this study, a comprehensive approach using molecular docking, in vitro assays, and molecular dynamics simulations was applied to identify potential inhibitors for SARS-CoV-2 papain-like protease (PLpro), a key and underexplored viral enzyme target. A focused protease inhibitor library was initially created and molecular docking was performed using CmDock software (v0.2.0), resulting in the selection of hit compounds for in vitro testing on the isolated enzyme. Among them, compound 372 exhibited promising inhibitory properties against PLpro, with an IC50 value of 82 ± 34 µM. The compound also displayed a new triazolopyrimidinyl scaffold not yet represented within protease inhibitors. Molecular dynamics simulations demonstrated the favorable binding properties of compound 372. Structural analysis highlighted its key interactions with PLpro, and we stress its potential for further optimization. Moreover, besides compound 372 as a candidate for PLpro inhibitor development, this study elaborates on the PLpro binding site dynamics and provides a valuable contribution for further efforts in pan-coronaviral PLpro inhibitor development.
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Dairy protein hydrolysates possess a broad spectrum of bioactivity and hypoallergenic properties, as well as pronounced bitter taste. The bitterness is reduced by complexing the proteolysis products with cyclodextrins (CDs), and it is also important to study the bioactivity of the peptides in inclusion complexes. Hydrolysates of whey and colostrum proteins with extensive hydrolysis degree and their complexes with ß/γ-CD were obtained in the present study, and comprehensive comparative analysis of the experimental samples was performed. The interaction of CD with peptides was confirmed via different methods. Bioactivity of the initial hydrolysates and their complexes were evaluated. Antioxidant activity (AOA) was determined by fluorescence reduction of fluorescein in the Fenton system. Antigenic properties were studied by competitive enzyme immunoassay. Antimutagenic effect was estimated in the Ames test. According to the experimental data, a 2.17/2.78-fold and 1.45/2.14-fold increase in the AOA was found in the ß/γ-CD interaction with whey and colostrum hydrolysates, respectively. A 5.6/5.3-fold decrease in the antigenicity of whey peptides in complex with ß/γ-CD was detected, while the antimutagenic effect in the host-guest systems was comparable to the initial hydrolysates. Thus, bioactive CD complexes with dairy peptides were obtained. Complexes are applicable as a component of specialized foods (sports, diet).
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Antimutagênicos , gama-Ciclodextrinas , Feminino , Gravidez , Humanos , Soro do Leite , Colostro , Proteínas do Soro do Leite/farmacologia , Peptídeos/farmacologiaRESUMO
Gold nanoparticles (AuNPs) are promising candidates in various biomedical applications such as sensors, imaging, and cancer therapy. Understanding the influence of AuNPs on lipid membranes is important to assure their safety in the biological environment and to improve their scope in nanomedicine. In this regard, the present study aimed to analyze the effects of different concentrations (0.5, 1, and 2 wt.%) of dodecanethiol functionalized hydrophobic AuNPs on the structure and fluidity of zwitterionic 1-stearoyl-2-oleoyl-sn-glycerol-3-phosphocholine (SOPC) lipid bilayer membranes using Fourier-transform infrared (FTIR) spectroscopy and fluorescent spectroscopy. The size of AuNPs was found to be 2.2 ± 1.1 nm using transmission electron microscopy. FTIR results have shown that the AuNPs induced a slight shift in methylene stretching bands, while the band positions of carbonyl and phosphate group stretching were unaffected. Temperature-dependent fluorescent anisotropy measurements showed that the incorporation of AuNPs up to 2 wt.% did not affect the lipid order in membranes. Overall, these results indicate that the hydrophobic AuNPs in the studied concentration did not cause any significant alterations in the structure and membrane fluidity, which suggests the suitability of these particles to form liposome-AuNP hybrids for diverse biomedical applications including drug delivery and therapy.
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Ouro , Nanopartículas Metálicas , Ouro/química , Nanopartículas Metálicas/química , Bicamadas Lipídicas/química , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Propolis is a resinous compound made by bees with well-known biological activity. However, comparisons between encapsulated and non-encapsulated propolis are lacking. Therefore, the antibacterial activity, effect on the phase transition of lipids, and inhibition of UV-induced lipid oxidation of the two forms of propolis were compared. The results showed that non-encapsulated propolis produces quicker effects, thus being better suited when more immediate effects are required (e.g., antibacterial activity). In order to gain an in-depth introspective on these effects, we further studied the synergistic effect of propolis compounds on the integrity of lipid membranes. The knowledge of component synergism is important for the understanding of effective propolis pathways and for the perspective of modes of action of synergism between different polyphenols in various extracts. Thus, five representative molecules, all previously isolated from propolis (chrysin, quercetin, trans-ferulic acid, caffeic acid, (-)-epigallocatechin-3-gallate) were mixed, and their synergistic effects on lipid bilayers were investigated, mainly using DSC. The results showed that some compounds (quercetin, chrysin) exhibit synergism, whereas others (caffeic acid, t-ferulic acid) do not show any such effects. The results also showed that the synergistic effects of mixtures composed from several different compounds are extremely complex to study, and that their prediction requires further modeling approaches.
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Própole , Própole/farmacologia , Quercetina/farmacologia , Flavonoides/farmacologia , Bactérias , Antibacterianos/farmacologia , LipídeosRESUMO
Extremophiles inhabit a wide variety of environments. Here we focus on extremophiles in moderate climates in central Europe, and particularly in Slovenia. Although multiple types of stress often occur in the same habitat, extremophiles are generally combined into groups according to the main stressor to which they are adapted. Several types of extremophiles, e.g., oligotrophs, are well represented and diverse in subsurface environments and karst regions. Psychrophiles thrive in ice caves and depressions with eternal snow and ice, with several globally distributed snow algae and psychrophilic bacteria that have been discovered in alpine glaciers. However, this area requires further research. Halophiles thrive in salterns while thermophiles inhabit thermal springs, although there is little data on such microorganisms in central Europe, despite many taxa being found globally. This review also includes the potential use of extremophiles in biotechnology and bioremediation applications.
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Lutein is a challenging compound to incorporate into food, as it is poorly soluble and unstable in aqueous solutions. In this study, the aim was to prepare stable encapsulates of lutein and lutein esters using feasible and straightforward techniques. Fine suspensions based on polyoxyethylene sorbitan monooleate and medium-chain triglyceride oil micelle-like units with 3.45% lutein esters or 1.9% lutein equivalents provided high encapsulation efficiencies of 79% and 83%, respectively. Lutein encapsulated in fine suspensions showed superior stability, as 86% was retained within the formulation over 250 days at 25 °C in the dark. Under the same storage conditions, only 38% of lutein remained in corresponding formulations. Higher encapsulation efficiencies were achieved with lecithin emulsions, at up to 99.3% for formulations with lutein, and up to 91.4% with lutein esters. In lecithin emulsions that were stored for 250 days, 17% and 80% of lutein and lutein esters, respectively, were retained within the formulations.
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Electrophilic aromatic substitution (EAS) is one of the most widely researched transforms in synthetic organic chemistry. Numerous studies have been carried out to provide an understanding of the nature of its reactivity pattern. There is now a need for a concise and general, but detailed and up-to-date, overview. The basic principles behind EAS are essential to our understanding of what the mechanisms underlying EAS are. To date, textbook overviews of EAS have provided little information about the mechanistic pathways and chemical species involved. In this review, the aim is to gather and present the up-to-date information relating to reactivity in EAS, with the implication that some of the key concepts will be discussed in a scientifically concise manner. In addition, the information presented herein suggests certain new possibilities to advance EAS theory, with particular emphasis on the role of modern instrumental and theoretical techniques in EAS reactivity monitoring.
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Knotweed is a flowering plant that is native to temperate and subtropical regions in the northern hemisphere. We evaluated Japanese (Reynoutria japonica Houtt.) and Bohemian (Fallopia x bohemica) knotweed rhizome and flower ethanol extracts and compared them in terms of their biological activities. The specific polyphenols were identified and quantified using HPLC/DAD, and the antioxidant activity was determined using 2,2-diphenly-1-picrylhydrazyl (DPPH) and cellular antioxidant capacity assays. The anticancer activity was evaluated as the difference between the cytotoxicity to cancer cells compared with control cells. The antimicrobial activity was determined using bacteria and yeast. The antidiabetic activity was tested as the ability of the extracts to inhibit α-amylase. Both rhizome extracts were sources of polyphenols, particularly polydatin and (-)-epicatechin; however, the cellular assay showed the highest antioxidant capacity in the flower extract of F. bohemica. The PaTu cell line was the least sensitive toward all knotweed extracts. The flower extracts of both species were less toxic than the rhizomes. However, the activity of the tested extracts was not specific for cancer cells, indicating a rather toxic mode of action. Furthermore, all used extracts decreased the α-amylase activity, and the rhizome extracts were more effective than the flower extracts. None of the extracts inhibited bacterial growth; however, they inhibited yeast growth. The results confirmed that rhizomes of Reynoutria japonica Houtt. could become a new source of bioactive compounds, which could be used for the co-treatment of diabetes and as antifungal agents.
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Neurodegenerative diseases, namely Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis, Huntington's disease, and multiple sclerosis are becoming one of the main health concerns due to the increasing aging of the world's population. These diseases often share the same biological mechanisms, including neuroinflammation, oxidative stress, and/or protein fibrillation. Recently, there have been many studies published pointing out the possibilities to reduce and postpone the clinical manifestation of these deadly diseases through lifelong consumption of some crucial dietary substances, among which phytochemicals (e.g., polyphenols) and endogenous substances (e.g., acetyl-L-carnitine, coenzyme Q10, n-3 poysaturated fatty acids) showed the most promising results. Another important issue that has been pointed out recently is the availability of these substances to the central nervous system, where they have to be present in high enough concentrations in order to exhibit their neuroprotective properties. As so, such the aim of this review is to summarize the recent findings regarding neuroprotective substances, their mechanisms of action, as well as to point out therapeutic considerations, including their bioavailability and safety for humans.
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Suplementos Nutricionais/normas , Doenças Neurodegenerativas/prevenção & controle , Ubiquinona/análogos & derivados , Humanos , Ubiquinona/farmacologia , Ubiquinona/uso terapêuticoRESUMO
: This study examined the hepatoprotective and anti-inflammatory effects of anthocyanins from Vaccinim myrtillus (bilberry) fruit extract on the acute liver failure caused by carbon tetrachloride-CCl4 (3 mL/kg, i.p.). The preventive treatment of the bilberry extract (200 mg anthocyanins/kg, orally, 7 days) prior to the exposure to the CCl4 resulted in an evident decrease in markers of liver damage (glutamate dehydrogenase, sorbitol dehydrogenase, malate dehydrogenase), and reduced pro-oxidative (conjugated dienes, lipid hydroperoxide, thiobarbituric acid reactive substances, advanced oxidation protein products, NADPH oxidase, hydrogen peroxide, oxidized glutathione), and pro-inflammatory markers (tumor necrosis factor-alpha, interleukin-6, nitrite, myeloperoxidase, inducible nitric oxide synthase, cyclooxygenase-2, CD68, lipocalin-2), and also caused a significant decrease in the dissipation of the liver antioxidative defence capacities (reduced glutathione, glutathione S-transferase, and quinone reductase) in comparison to the results detected in the animals treated with CCl4 exclusively. The administration of the anthocyanins prevented the arginine metabolism's diversion towards the citrulline, decreased the catabolism of polyamines (the activity of putrescine oxidase and spermine oxidase), and significantly reduced the excessive activation and hyperplasia of the Kupffer cells. There was also an absence of necrosis, in regard to the toxic effect of CCl4 alone. The hepatoprotective mechanisms of bilberry extract are based on the inhibition of pro-oxidative mediators, strong anti-inflammatory properties, inducing of hepatic phase II antioxidant enzymes (glutathione S-transferase, quinone reductase) and reduced glutathione, hypoplasia of Kupffer cells, and a decrease in the catabolism of polyamines.
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Multifunctional liposomes incorporating ß-sitosterol were developed for delivery of gentisic acid (GA). The interactions of both compounds with phospholipid bilayer were interpreted viaeffects of different ß-sitosterol content (0, 20 and 50â¯mol %) and different gentisic acid to lipid ratio (nGA/nlip from 10-5 to 1) on membrane fluidity and thermotropic properties. Multilamellar vesicles of phosphatidylcholines (with size range between 1350 and 1900â¯nm) effectively encapsulated GA (54%) when nGA/nlip was higher than 0.01. Suppression of lipid peroxidation was directly related to concentration of GA. The resistance to diffusion of gentisic acid from liposomes increased for Ë50% in samples incorporating 50â¯mol % ß-sitosterol compared to sterol-free liposomes. Finally, simulated in vitro gastrointestinal conditions showed that the release was mainly affected by low pH of simulated gastric fluid and the presence of cholates in simulated intestinal fluid, rather than by enzymes activity.
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Gentisatos/química , Bicamadas Lipídicas/química , Lipossomos/química , Fosfatidilcolinas/química , Sitosteroides/metabolismo , Materiais Biomiméticos/química , Compostos de Boro/química , Difusão , Composição de Medicamentos/métodos , Liberação Controlada de Fármacos , Corantes Fluorescentes/química , Suco Gástrico/química , Gentisatos/farmacologia , Concentração de Íons de Hidrogênio , Cinética , Peroxidação de Lipídeos/efeitos dos fármacos , Fluidez de Membrana/efeitos dos fármacos , Sitosteroides/química , Relação Estrutura-AtividadeRESUMO
The present paper reports a novel method to improve the properties of polyethylene (PE) and polypropylene (PP) polymer foils suitable for applications in food packaging. It relates to the adsorption of chitosan-colloidal systems onto untreated and oxygen plasma-treated foil surfaces. It is hypothesized that the first coated layer of chitosan macromolecular solution enables excellent antibacterial properties, while the second (uppermost) layer contains a network of polyphenol resveratrol, embedded into chitosan nanoparticles, which enables antioxidant and antimicrobial properties simultaneously. X-ray photon spectroscopy (XPS) and infrared spectroscopy (FTIR) showed successful binding of both coatings onto foils as confirmed by gravimetric method. In addition, both attached layers (chitosan macromolecular solution and dispersion of chitosan nanoparticles with incorporated resveratrol) onto foils reduced oxygen permeability and wetting contact angle of foils; the latter indicates good anti-fog foil properties. Reduction of both oxygen permeability and wetting contact angle is more pronounced when foils are previously activated by O2 plasma. Moreover, oxygen plasma treatment improves stability and adhesion of chitosan structured adsorbates onto PP and PE foils. Foils also exhibit over 90% reduction of Staphylococcus aureus and over 77% reduction of Escherichia coli as compared to untreated foils and increase antioxidant activity for over a factor of 10. The present method may be useful in different packaging applications such as food (meat, vegetables, dairy, and bakery products) and pharmaceutical packaging, where such properties of foils are desired.
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We studied seven known peptides derived from different food proteins, and one hydrophobic peptide derived through in-silico hydrolysis. We investigated their antioxidative, antimicrobial and haemolytic activities and their interactions with model lipid membranes. Our data reveal that peptides with histidines are the most effective for protection against lipid peroxidation, as: P#3 (VHHA)â¯>â¯P#5 (LHALLL)â¯>â¯P#7 (LLPHH). The most potent peptide towards metal-induced oxidation was P#2 (LQKW), which contained an aromatic tryptophan at its C-terminus. The peptides with negative hydrophobicity, as P#1 (PEL) and P#2 (LQKW), were the most potent according to DPPH radical scavenging. The most potent peptide towards superoxide radicals was the four-amino-acid chain with aromatic amino acids, P#2 (LQKW). The peptides studied did not show haemolytic or antimicrobial activities, except P#4 (AAGGV) against Listeria monocytogenes. These peptides did not interact with model lipid membranes, except P#8, which was designed to span lipid bilayers.
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Antioxidantes/farmacologia , Lipídeos de Membrana/química , Peptídeos/química , Peptídeos/farmacologia , Animais , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Antioxidantes/química , Varredura Diferencial de Calorimetria , Bovinos , Avaliação Pré-Clínica de Medicamentos/métodos , Hemólise/efeitos dos fármacos , Interações Hidrofóbicas e Hidrofílicas , Bicamadas Lipídicas , Peroxidação de Lipídeos/efeitos dos fármacos , Listeria monocytogenes/efeitos dos fármacos , Lipídeos de Membrana/metabolismo , Peptídeos/isolamento & purificação , Espécies Reativas de Oxigênio/metabolismo , Espectrometria de Fluorescência , Relação Estrutura-Atividade , Superóxidos/química , Triptofano/químicaRESUMO
This study examined the nephroprotective effects of 15 different anthocyanins from the bilberry extract on the acute kidney injury caused by CCl4. The acute nephrotoxicity in rats was induced 24â¯h after the treatment with a single dose of CCl4 (3â¯mL/kg, i.p.).The nephroprotective effects of the anthocyanins were examined in the animals that had been given the bilberry extract in a single dose of 200â¯mg of anthocyanins/kg daily, 7 days orally, while on the seventh day, 3â¯h after the last dose of anthocyanins, the animals received a single dose of CCl4 (3â¯mL/kg, i.p.) and were sacrificed 24â¯h later. When the nephrotoxicant alone was administered, it resulted in a substantial increase of the pro-oxidative (TBARS, CD, H2O2, XO, and GSSG) and pro-inflammatory markers (TNF-α, NO, and MPO), as well as a noticeable reduction of the antioxidant enzymes (CAT, SOD, POD, GPx, GST, GR) and GSH when compared to the results of the control group. Moreover, the application of CCl4 significantly influenced a reduction of the renal function, as well as an increase in the sensitive and specific injury indicators of the kidney epithelial cells (ß2-microglobulin, NGAL, KIM1/TIM1) in the serum and urine of rats. The pretreatment of the animals poisoned with CCl4 with the anthocyanins from the bilberry extract led to a noticeable reduction in the pro-oxidative and pro-inflammatory markers with reduced consumption of the antioxidant defence kidney capacity, compared to the animals exposed to CCl4 alone. Anthocyanins have been protective for the kidney parenchyma, with an apparent absence of the tubular and periglomerular necrosis, severe degenerative changes, inflammatory mononuclear infiltrates and dilatation of proximal and distal tubules, in contrast to the CCl4-intoxicated animals. The nephroprotective effects of anthocyanins can be explained by strong antioxidant and anti-inflammatory effects achieved through the stabilization and neutralization of highly reactive and unstable toxic CCl4 metabolites.
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Antocianinas/farmacologia , Nefropatias/prevenção & controle , Vaccinium myrtillus/química , Animais , Antocianinas/química , Antocianinas/isolamento & purificação , Tetracloreto de Carbono , Nefropatias/induzido quimicamente , Nefropatias/metabolismo , Masculino , Extratos Vegetais , Ratos , Ratos WistarRESUMO
Herein, the protein binding rates of structurally different flavonoids to human serum albumin (HSA) were elucidated by applying the high performance affinity chromatography (HPAC). The flavonoids with hydroxyl groups on ring A showed a higher protein binding rate compared with those that there was no hydroxyl on ring A. However, the hydroxylation of ring B lowered the protein binding rate. It was also found that an additional methoxy group in flavone ring A would decrease the protein binding rate. Nevertheless, the methoxy group in flavanone ring A (position 6) and isoflavone ring B (position 4') increased the protein binding rate. Methoxy group at other positions of flavonoids slightly enhanced or no significantly affected the binding rates on human serum albumin. Hydrogenation of C2C3 double bond of flavonoids decreased the protein binding rate and had the same effect as glycosylation which decrease the protein binding rate by 5%-25%.
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Proteínas Sanguíneas/química , Cromatografia de Afinidade/métodos , Polifenóis/química , Albumina Sérica Humana/química , Flavonoides , Humanos , Ligação Proteica , Relação Estrutura-AtividadeRESUMO
Measuring and quantifying thermodynamic parameters that determine both the stability of and interactions between biological macromolecules are an essential and necessary complement to structural studies. Although basic thermodynamic parameters for an observed process can be readily obtained, the data interpretation is often slow and analysis quality can be extremely variable. We have started to develop a web application that will help users to perform thermodynamic characterizations of oligonucleotide unfolding. The application can perform global fitting of calorimetric and spectroscopic data, and uses a three-state equilibrium model to obtain thermodynamic parameters for each transition step - namely, the Gibbs energy, the enthalpy, and the heat capacity. In addition, the application can define the number of K+ ions and the number of water molecules being released or taken up during unfolding. To test our application, we used UV spectroscopy, circular dichroism, and differential scanning calorimetry to monitor folding and unfolding of a model 22-nucleotide-long sequence of a human 3'-telomeric overhang, known as Tel22. The obtained data were uploaded to the web application and the global fit revealed that unfolding of Tel22 involves at least one intermediate state, and that K+ ions are released during the unfolding, whereas water molecules are taken up.
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Archaeosomes are vesicles made of lipids from archaea. They possess many unique features in comparison to other lipid systems, with their high stability being the most prominent one, making them a promising system for biotechnological applications. Here, we report a preparation protocol of large unilamellar vesicles, giant unilamellar vesicles (GUVs), and nanodiscs from archaeal lipids with incorporated cholesterol. Incorporation of cholesterol led to additional increase in thermal stability of vesicles. Surface plasmon resonance, sedimentation assays, intrinsic tryptophan fluorescence measurements, calcein release experiments, and GUVs experiments showed that members of cholesterol-dependent cytolysins, listeriolysin O (LLO), and perfringolysin O (PFO), bind to cholesterol-rich archaeosomes and thereby retain their pore-forming activity. Interestingly, we observed specific binding of LLO, but not PFO, to archaeosomes even in the absence of cholesterol. This suggests a new capacity of LLO to bind to carbohydrate headgroups of archaeal lipids. Furthermore, we were able to express LLO inside GUVs by cell-free expression. GUVs made from archaeal lipids were highly stable, which could be beneficial for synthetic biology applications. In summary, our results describe novel model membrane systems for studying membrane interactions of proteins and their potential use in biotechnology.
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Archaea/metabolismo , Colesterol/química , Citotoxinas/química , Lipossomas Unilamelares/química , Colesterol/metabolismo , Citotoxinas/metabolismo , Lipossomas Unilamelares/metabolismoRESUMO
This study characterises the genetic variability of local pomegranate (Punica granatum L.) germplasm from the Slovenian and Croatian areas of Istria. The bioactive components and antioxidant and antimicrobial properties of ethanol and water extracts of different parts of pomegranate fruit were also determined, along with their preliminary nutritional characterisation. Twenty-six different genotypes identified with microsatellite analysis indicate the great diversity of pomegranate in Istria. The pomegranate fruit ethanol extracts represent rich sources of phenolic compounds (mean value of the mass fraction in exocarp and mesocarp expressed as gallic acid is 23 and 16 mg/g, respectively). The ethanol extracts of pomegranate exocarp and mesocarp showed the greatest antimicrobial activity against Candida albicans, Candida parapsilosis, Rhodotorula mucilaginosa, Exophiala dermatitidis and Staphylococcus aureus, and the same water extracts against S. aureus and Escherichia coli. To the best of our knowledge, this study represents the first report of the characterisation of pomegranate genetic resources from Istria at different levels, including the molecular, chemical, antimicrobial and nutritional properties.