Evaluation of the Effect of Antibacterial Peptides on Model Monolayers.

The aim of the study was to assess the effect of the synthesized antibacterial peptides: P2 (WKWK)2-KWKWK-NH2, P4 (C12)2-KKKK-NH2, P5 (KWK)2-KWWW-NH2, and P6 (KK)2-KWWW-NH2 on the physicochemical properties of a model biological membrane made of azolectin or lecithin. The Langmuir Wilhelmy method was used for the experiments. Based on the compressibility factor, it was determined that the monolayers formed of azolectin and peptides in the aqueous subphase are in the condensed liquid phase. At the boundary between the condensed and expanded liquid phases, there was a monolayer made of lecithin and P4, P5 or P6 in the aqueous subphase. In turn, the film consisting of lecithin alone (37.7 mN/m) and lecithin and P2 (42.6 mN/m) in the water subphase was in the expanded liquid phase. All peptides change, to varying degrees, the organization and packing of molecules in the monolayer, both those made of azolectin and of lecithin. The test results can be used for further research to design a system with the expected properties for specific organisms.

Effect of Newly Synthesized Structures of Peptides on the Stability of the Monolayers Formed.

The aim of the study was to evaluate the effect of the peptide structure (WKWK)2-KWKWK-NH2, P4 (C12)2-KKKK-NH2, P5 (KWK)2-KWWW-NH2, P6 (KK)2-KWWW-NH2 on their physicochemical properties. The thermogravimetric method (TG/DTG) was used, which made it possible to observe the course of chemical reactions and phase transformations occurring during the heating of solid samples. Based on the DSC curves, the enthalpy of the processes occurring in the peptides was determined. The influence of the chemical structure of this group of compounds on their film-forming properties was determined using the Langmuir-Wilhelmy trough method and was followed by molecular dynamics simulation. Evaluated peptides showed high thermal stability and the first significant mass loss occurred only at about 230 °C and 350 °C. The analysis of the compressibility coefficient of individual peptides indicates that all formed peptide monolayers were in the expanded liquid phase. Their maximum compressibility factor was less than 50.0 mN/m. Its highest value of 42.7 mN/m was achieved in a monolayer made of P4. The results obtained in molecular dynamic simulation indicate that non-polar side chains played an important role in the properties of the P4 monolayer, and the same applies to P5, except that a spherical effect was observed here. A slightly different behavior was observed for the P6 and P2 peptide systems, where the type of amino acids present had an influence. The obtained results indicate that the structure of the peptide affected its physicochemical and layer-forming properties.

Assessment of the Influence of the Selected Range of Visible Light Radiation on the Durability of the Gel with Ascorbic Acid and Its Derivative.

(1) Background: Depending on the type of hydrophilic polymer used, different types of hydrogels may be chemically stable or may degrade and eventually disintegrate, or dissolve upon exposure to sunlight. Many over-the-counter medications are now stored with a limited control of temperature, humidity and lighting. Therefore, in this study, the photostability of a gel made of cross-linked polyacrylic acid (PA), methylcellulose (MC) and aristoflex (AV) was assessed, and the interaction between the polymers used and ascorbic acid and its ethylated derivative was investigated. (2) Methods: The samples were continuously irradiated at constant temperature for six hours. The stability of the substance incorporated into the gels was assessed using a UV-Vis spectrophotometer. FTIR-ATR infrared spectroscopy was used to measure changes during the exposure. (3) Results: Ascorbic acid completely decomposed between the first and second hours of illumination in all samples. The exception is the preparation based on polyacrylic acid with glycerol, in which the decomposition of ascorbic acid slowed down significantly. After six hours of irradiation, the ethylated ascorbic acid derivative decomposed in about 5% for the polyacrylic acid-based gels and aristoflex, and in the methylcellulose gel it decomposed to about 2%. In the case of ascorbic acid, the most stable formulation was a gel based on polyacrylic acid and polyacrylic acid with glycerol, and in the case of the ethyl derivative, a gel based on methylcellulose. (4) Conclusions: The experiment showed significant differences in the decomposition rate of both compounds, resulting from their photostability and the polymer used in the hydrogel.

Antimicrobial and Antioxidative Activity of Newly Synthesized Peptides Absorbed into Bacterial Cellulose Carrier against Acne vulgaris.

The ongoing search for effective treatment of Acne vulgaris is concentrated, i.a., on natural peptides with antimicrobial properties. The aim of this work was the development of new amino acid derivatives with potential activity on dermal infections against selected microorganisms, including the facultative anaerobe C. acne. The peptides P1-P6 were synthesized via Fmoc solid phase peptide synthesis using Rink amide AM resin, analyzed by RP-HPLC-MS, FTIR, DPPH radical scavenging activity, and evaluated against C. acne and S. aureus, both deposited and non-deposited in BC. Peptides P1-P6 presented a lack of cytotoxicity, antimicrobial activity, or antioxidative properties correlated with selected structural properties. P2 and P4-P6 sorption in BC resulted in variable data, i.a., confirming the prospective topical application of these peptides in a BC carrier.

The Influence of UV Radiation on the Degradation of Pharmaceutical Formulations Containing Quercetin.

The aim of this study was to assess the photostability of quercetin in the presence of anionic and nonionic polymeric gels with varied compositions of an added component-glycerol. The samples were irradiated continuously at constant temperature. The stability of quercetin in solution and incorporated into the gels was evaluated by an UV-Vis spectrophotometer. FTIR spectroscopy (Fourier-transform infrared spectroscopy) was used to detect the changes in the structure of quercetin depending on the polymer used in the gel, and on the exposure time. Photostabilization is an important aspect of quality assurance in photosensitive compounds. The decomposition rate of quercetin in the ionic preparation of polyacrylic acid (PAA) with glycerol was 1.952·10-3 min-1, whereas the absence of glycerol resulted in a decay rate of 5.032·10-4 min-1. The formulation containing non-ionic methylcellulose resulted in a decomposition rate of quercetin in the range of 1.679·10-3 min-1. The decay rate of quercetin under light influence depended on the composition of the gel. It was found that the cross-linked PAA stabilized quercetin and the addition of glycerol accelerated the photodegradation.

Edible alginate/chitosan-based nanocomposite microspheres as delivery vehicles of omega-3 rich oils.

A new series of alginate/chitosan-based nanocomposite microspheres was developed to achieve the maximum health benefit and to minimize the oxidation of omega-3 rich oils (flaxseed or fish oils). The nanocomposite microspheres incorporate curcumin (Cur) as natural antioxidant, and have been prepared using a three-step procedure (oil-in-water (o/w) emulsification, gelation and microencapsulation). The average particle size of Cur-free and Cur-loaded nanocomposites ranged between 139 and 153 nm. The presence of omega-3 rich oils in core of the formulated microspheres was confirmed by XRD and FTIR. Optical microscopy, stereo microscopy, SEM and AFM showed a spherical shape of the microspheres. Microencapsulation efficiency, oxidative stability, release profile of oils as well as the antioxidant and antibacterial activities were investigated. The results suggested that the microspheres could be applied as effective and safe edible vehicles for hydrophobic nutraceuticals like omega-3 rich oils with broad spectrum antibacterial activity.

Antioxidant and antibacterial activities of omega-3 rich oils/curcumin nanoemulsions loaded in chitosan and alginate-based microbeads.

Omega-3 fatty acids can be considered as potential alternative therapeutic agents because of their antimicrobial and antioxidant properties. Two investigated omega-3 rich oils (flaxseed or fish) have been nanoemulsified with and without the natural antioxidant (curcumin, Cur) followed by their incorporation into crosslinked polymeric microbeads. The microbeads were developed from chitosan (CS), alginate (AL) and their combination (CSAL). Results indicated that the mean droplet diameter of the plain and Cur-loaded nanoemulsions ranged from 62.3 to 111.29 nm. The microbeads produced from AL, CS and their combination without Cur had predominantly shriveled surfaces compared to Cur-loaded ones. Addition of Cur was found to enhance oxidative stability, encapsulation efficiency, loading capacity, and antioxidant activity of the formulated microbeads. Plain fish oil revealed more antibacterial activity than plain flaxseed oil. Fish oil nanoemulsion-in-AL microbeads had more antibacterial activity than nanoemulsions of flaxseed oil-in-AL, fish oil-in-CS and the combined (CSAL) microbeads. However, flaxseed oil nanoemulsion-in-CS microbeads showed higher antibacterial activity than nanoemulsions of fish oil-in-CS, flaxseed oil-in-AL and the combined microbeads. The obtained results suggested the suitability of the formulated nanoemulsions-loaded microbeads to be used in food and pharmaceuticals products.

Selected Physicochemical and Biological Properties of Ethyl Ascorbic Acid Compared to Ascorbic Acid.

The aim of the work was the evaluation of selected biological and physicochemical, applicative properties of ethylated ascorbic acid (AAE) compared to ascorbic acid (AA). Thermogravimetry (TG), differential thermogravimetry (DTG), and differential thermal analysis (DTA) were conducted, followed by the evaluation of AAE decomposition by the UV-Vis spectroscopic method including the influence of temperature and pH. Scavenging, antimicrobial activity and cytotoxicity against L929 fibroblasts were also performed. The difference in mass loss between AA and AAE was 30% via TG. DTA revealed characteristic exothermic and endothermic effects. The AAE solution was more thermally stable than AA. The calculated zero-order rate constants of free-radical scavenging kinetics for AAE were in the range of 4.9×10-3-1.35×10-2 s-1. The activation energy for the process was 11,2281 kJ/mol. AAE was active against Staphylococcus (S.) aureus and Enterococcus (E.) faecalis and acted stronger against Candida (C.) albicans than AA. The concentrations of AA ≥2.5% were cytotoxic, whereas in the case of AAE, a 10% concentration was considered cytotoxic. DTG enables the detailed differentiation between AA and AAE. AAE in aqueous solution is more stable compared to AA. The antioxidant activity of AAE is significant. However, the reaction with 2,2-diphenyl-1-picrylhydrazyl (DPPH) indicates prolonged activity compared to AA. Variability in the antimicrobial activity of AAE may find practical application in the pharmaceutical and cosmetic industries. The potential for applicative aims may be supported by the relatively low in vitro toxicity of AAE.

Application of an anionic polymer in the formulation of floating tablets containing an alkaline model drug.

BACKGROUND: Gastric residence time is the key factor affecting the bioavailability of active pharmaceutical ingredients absorbed mainly through the gastric mucous membrane and influencing the local activity of some drugs. OBJECTIVES: The aim of this study was the development of a new composition of non-effervescent floating tablets and the evaluation of the effect of an anionic polymer and compressive force on the floating properties and release characteristics of tablets containing a model alkaline drug, chlorhexidine (CHX). MATERIAL AND METHODS: Direct compression was applied to a polyacrylic acid derivative and sorbitol to fabricate the tablets. Drug release was analyzed using several kinetic models. The formulations floated on the surface of the fluid for 24 h. The values of the rate constants, statistical parameters, and half-release time (t0.5) were calculated. RESULTS: The diffusion coefficient n falls between 0.54 ±0.02 and 0.81 ±0.03 for most formulations. The floating time (FT) and floating lag time (FLT) were found to depend on the amount of polymer incorporated in the formulations. A high compressive force sustained the release of the drug but reduced the FT and FLT. Based on the FT and t0.5, it was determined that the C1 composition is the optimal formulation with FT >24 h and t0.5 between 113 ±2 and 144 ±13 min, depending on the drug release model. CONCLUSIONS: The application of an anionic polymer results in a prolonged release of the drug from the tablets and allows them to float on fluid surfaces.

Antioxidant activity of selected phenols estimated by ABTS and FRAP methods.

INTRODUCTION: Phenols are the most abundant compounds in nature. They are strong antioxidants. Too high level of free radicals leads to cell and tissue damage, which may cause asthma, Alzheimer disease, cancers, etc. Taking phenolics with the diet as supplements or natural medicines is important for homeostasis of the organism. MATERIALS AND METHODS: The ten most popular water soluble phenols were chosen for the experiment to investigate their antioxidant properties using ABTS radical scavenging capacity assay and ferric reducing antioxidant potential (FRAP) assay. RESULTS AND DISCUSSION: Antioxidant properties of selected phenols in the ABTS test expressed as IC50 ranged from 4.332 µM to 852.713 µM (for gallic acid and 4- hydroxyphenylacetic acid respectively). Antioxidant properties in the FRAP test are expressed as µmol Fe2+/ml. All examined phenols reduced ferric ions at concentration 1.00 x 10-3 mg/ml. Both methods are very useful for determination of antioxidant capacity of water soluble phenols.

Organic matter transformation in the environment investigated by quantitative electron paramagnetic resonance (EPR) spectroscopy: studies on lignins.

The lignins separated from angiosperm and gymnosperm trees, peat and xylitic brown coal were investigated by quantitative EPR. Observed free radicals in lignins are sensitive to alkaline environment. Gaseous ammonia interacting with solid lignins in resonance cavity shifts quinone-hydroquinone equilibria towards formation of semiquinone anions. Complexation of copper(II) by lignins causes drastic decrease of the semiquinones in the matrices. Formation of lignin-Pb(II) complexes yielded radicals characterised by unusually low g-value (1.9999-2.0003). Monomeric structural units of the investigated lignins were recognised by pyrolysis with in situ methylation by tetramethylammonium hydroxide. Although for the natural lignins the mixture of normal semiquinone signals at g about 2.0034 and signals at g 1.9999 were observed, some monomeric components of lignins (e.g., caffeic acid, pyrogallol) gave pure lines at g = 1.9999. The bacterial oxidative biodegradation of lignin monomeric components and their Pb(II) complexes resulted in increase of the radical signals.

Quantitative EPR study on free radicals in the natural polyphenols interacting with metal ions and other environmental pollutants.

Quantitative electron paramagnetic resonance (EPR) method was applied to characterise radicals stabilised in polyphenolic matrices of various biogenic materials: lichens, mosses, composts, soils, peats, brown coals and sewage sludge sediments. The investigations were carried out on raw materials and extracted fractions of humic acids (HA). General trends of g value and spin concentration changes were found. These parameters in lichens strongly depend on lichen species and air pollution. Determination of the g value and semiquinone spin concentration allows to assess degree of transformation of organic matter in compost, soil, peat and lignite. Application of gaseous ammonia as a base penetrating the organic matrices extends the possibilities and usefulness of the method. Interaction of metal ions with humic materials is illustrated by interaction of VO2+ ion with peat and lignite HA as well as demineralised (raw and carbonised) brown coal. Our investigations demonstrate that quantitative EPR is a rapid and effective monitoring method to study the influence of various environmental factors on substances containing polyphenolic matrices.