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This paper is focused on the development of the experimental environment connected to reactor graphite. Regarding its very good neutronic and mechanical properties, graphite will be very important in some new reactor designs, such as high-temperature or molten salt SMR reactors. These new reactor concepts require a new experimental environment as support for further research. In the laboratories of the Research Centre Řež and at the LR-0 reactor, the new experimentally validated graphite environment was created. This large graphite insertion is the largest graphite mono-block, which is possible to assemble at the LR-0 reactor. Sets of experiments for measuring reaction rates of different activation detectors for neutron field mapping were performed. This approach was used for thermal and epithermal region descriptions. For the fast neutron spectrum evaluation, the stilbene scintillation detector was used. All parameters, such as criticality height of moderator level, neutron spectrum, and other parameters for all experiments, were performed using Monte Carlo neutronic codes Serpent and MCNP. The obtained results were finally compared to the measurement of neutron leakage spectra from the graphite cube and graphite cylinder. These specially developed graphite-shaped neutron fields, reactor insertions, and external cube and cylinder with Cf neutron source can be used in the future for validation of not only materials used in SMR reactors but for arbitrary cross-section verification.

Red yeast Cystofilobasidium capitatum autofluorescence was studied by means of confocal laser scanning microscopy (CLSM) to reveal distribution of carotenoids inside the cells. Yeasts were cultivated in 2L fermentor on glucose medium at permanent light exposure and aeration. Samples were collected at different times for CLSM, gravimetric determination of biomass and HPLC determination of pigments. To compare FLIM (Fluorescence Lifetime Imaging Microscopy) images and coupled data (obtained by CLSM) with model systems, FLIM analysis was performed on micelles of SDS:ergosterol and SDS:coenzyme Q with different content of ergosterol and coenzyme Q, respectively, and with constant addition of beta-carotene. Liposomes lecithin:ergosterol:beta-carotene were investigated too. Two different intracellular forms of carotenoids were observed during most of cultivations, with third form appeared at the beginning of stationary phase. Observed behavior is probably due to formation of some kind of carotenoid protective system in membranes of different compartments of yeast cell, especially cytoplasmic membrane.

This article focuses on description of two different neutron fields from linear and cylindrical Inertial Electrostatic Confinement (IEC) neutron generators. Both of these generators are well defined and commonly used. They use a deuterium-tritium reaction that produces neutrons with energies in the range 13 – 16 MeV, depending on the direction and the energy of the incoming deuterium nucleus. Two-parametric spectrometric system for neutron/gamma mixed fields NGA-01 was used to characterize neutron spectra in the proximity of generators. The cylindrical 45x45 mm stilbene scintillator was connected to this device using an active voltage divider. This way, we were able to measure neutron energies in the range 1 - 15 MeV while filtering out gamma radiation, even when counts per second is high. For the neutron spectrum calculation recoil spectra using deconvolution through maximum likelihood estimation was used. Measured neutron spectra have been compared with simulations realized via MCNP6. According to the theoretical prediction, these two types of generators produce different neutron fields. In case of the linear generator the target is very close to point located tritium bombarded by deuterons. Thus the neutron spectrum varies depending on the angle between the detector axis and the axis of the generator. Both experimental results and simulation show a shift of the neutron energy peak in pulse height histogram. For IEC type generators the neutron spectrum is more complicated. The shape and the position of the neutron energy peak depend heavily on the position of the detector. The most prominent effect is in the position in the plane perpendicular to the generator axis. In this case, the peak splits into two peaks that can be measured and distinguished. These results were verified by the diamond detector which was also used for characterization of the IEC type generator.

Pool-type reactors are an excellent source of high energy photons without neutrons because the water above fuel acts as an outstanding neutron shielding, whereas for high energy photons, it is nearly transparent. The absence of neutrons is a valuable property essential for testing semiconductor detectors because they are sensitive to neutrons, which might cause their damage. The high energy gammas are worth studying because they are responsible for radiation induced heating of reactor internal components in energy producing reactors, which might lead to their radiation induced damage by void swelling. At this effect, the helium produced in (n,α) reactions influenced by locally increased temperature clusters into bigger bubbles, which are increasing material volume, and therefore changing the geometry of internals. The referred method could be utilized e.g. for resource exploration by detecting prompt capture gammas. The prompt gamma field was measured at the VR-1 reactor with two different cores and water levels. The stilbene measurements showed relatively good agreement with the spectrum shape computed by MCNP6.2. An important result is a confirmation that the stilbene deconvolution matrix can be used with a rotated crystal. The testing measurement with HPGe detectors showed a possibility for measurement in such type of geometry.

In this paper we study quality of three types of organic scintillators stilbene, p-terphenyl and EJ-299-33. We used monoenergetic neutron fields with a wide range of neutron energies in PTB Braunschweig. All the tests were carried out with NGA-01 spectrometer. The results of the measurements are evaluated spectra from the spectrometer. We discuss the quality of each scintillator, such as the FWHM of the peaks.

Liquid organic scintillators are important devices for measurements of neutron radiation. This work aims to develop and optimize the composition of liquid organic scintillators so it can be used for fast neutron spectrometry. As the neutron radiation is usually accompanied with γ ray radiation, it is important to have quality γ/n discrimination. The new cocktail for house made liquid organic scintillator is prepared and studied with intention of being able to separate gamma and neutron for neutron energies above 0.5 MeV while keeping lower constraints on practical use (e.g., sealing because of oxygen) than commercial liquid scintillators. In preceding work the composition of liquid scintillators was optimized. Two twocomponent scintillators were selected for further studies. Solvent DIPN (Di-iso-propyl-naphthalene Mixed Isomers) is selected for both. First is mixed with luminophore PYR (1-Phenyl-3-(2,4,6-trimethyl-phenyl)-2-pyrazoline) of concentration 5 g/l. Second is mixed with luminophore THIO (2,5-Bis(5-tert-butyl-benzoxazol-2-yl)thiophene) of concentration 5 g/l. In this work the response of scintillator to monoenergetic beam of neutrons was measured for multiple neutron energies at PTB in Braunschweig. The two parameter spectrometric system NGA-01 is used to analyze the energy and discrimination characteristics. 137 Cs and 60 Co are used as radiation sources for calibration with pure γ rays. Then the response of scintillator for same neutron energies was simulated using GEANT4. The dissipated energy in the scintillator in response to monoenergetic neutrons is obtained. Both, measured and simulated responses are compared. Functional dependence for yield of recoiled products is estimated. It is seen that main recoil product hydrogen proton is well observed in both. From the edge of proton response one can assume the yield for given neutron energy. The recoiled carbon ion (from elastic collision) is on the other side difficult to observe in measured results but clearly seen in dissipated energy plots. It suggests that yield of carbon ion is very small relatively to proton yield. These results will serve as basis for response function evaluation of scintillator which is necessary for evaluation of unknown neutron spectra from measurements with scintillator.

Physical quantities derived from integral experiments can usually be measured much more accurately than that from differential nuclear data. The accurate knowledge of integral parameters provide excellent grounds for testing and tuning differential data such as, for instance, cross sections. Measurement of neutron leakage spectra with 252Cf neutron source located at sphere center is often used for integral experiments. While this type of experiments provide information for cross section tuning, however, care must be taken to avoid misleading interpretation, namely, at high energies due to the very low portion of high energy neutrons in 252Cf spectrum. This issue can be alleviated by the use of point source with different spectra shape. For that purpose one suitable candidate seems to be the AmBe neutron source which has a relatively high average energy and peak character of emitted neutrons. Indeed, AmBe seems an interesting option because the calculated leakage neutron spectra are not very sensitive to the input shape of the neutron spectra. Thus the neutron leakage spectra calculated using tabulated of International Organization for Standardization spectra is nearly the same as stilbene measured AmBe spectra as an input.

This paper presents some results of the development of two new scintillation detectors usable for the detection and spectrometry of neutron and photon components of the mixed field. It is a series of liquid organic scintillators containing carbon, hydrogen, nitrogen and oxygen. Elements of individual scintillator designs are arranged in various chemical bonds defining physical and chemical properties of the scintillators. These scintillators have advantageous useful properties. They are non-hygroscopic, not poisonous and are not sensitive to contact with the outside atmosphere and can be stored in conventional laboratory containers.

This paper has been formally retracted because it has been accidentally published twice in the same volume. Request approved by the proceedings Editor and the Publisher on June 02, 2023.

In this work we compare the pulse-shape discrimination (PSD) properties of EJ-299-33A, BC-501A, stilbene, p-terphenyl and Hidex Aqualight in neutron field generated by the LVR-15 reactor with silicon filter utilization. Pulses from the scintillators are processed by Neutron-Gamma spectrometer. This spectrometric system with fast digitizer card contains two analog-digital (A/D) converters with a resolution of 12 bits and sampling frequency 500 MHz. For photomultiplier (PMT) linearity improvement active divider has been used. Measured data from scintillators have been processed using the integration method and compared. Results are presented.

This paper presents some results of the development of two new scintillation detectors usable for the detection and spectrometry of neutron and photon components of the mixed field. It is a series of liquid organic scintillators containing carbon, hydrogen, nitrogen and oxygen. Elements of individual scintillator designs are arranged in various chemical bonds defining physical and chemical properties of the scintillators. These scintillators have advantageous useful properties. They are non-hygroscopic, not poisonous and are not sensitive to contact with the outside atmosphere and can be stored in conventional laboratory containers.

LR-0 reactor is an experimental reactor in NRI Rez, Czech Republic. So far an analog apparatus was used for measurements of the space-energy distribution of the neutron gamma mixed field inside the reactor vessel. Recently we measured in LR-0 with fully digital apparatus using Agilent digitizer and compared our results with older established results from analog apparatus and also with MCNP calculations.

Complex flow structures emerging in human lungs significantly affect deposition of inhaled particles. Therefore flow measurements alongside with deposition measurements have to be performed on the same model geometry. Amount of deposited aerosol particles in different regions of lungs could be measured by several methods. Gamma detection of radioactive tagged particles is subjected to strict regulations and is expensive; gravimetry and fluorometry are thus preferred in current studies. Novel methodology for fluorometric measurement of aerosol transport in models of human lungs was developed. Some previously published experiments were performed using condensation of di(2-ethylhexyl) sebacate (DEHS) vapours on fluorescein nuclei. Their authors assumed that fluorescein is necessary for generation of fluorescent particles and they neglected possible fluorescence of DEHS itself. Our analysis of DEHS characteristics indicated that DEHS fluorescence could be detected apart in sufficient rate to evaluate aerosol deposition. Consequently experiments in standard setup of condensation monodisperse aerosol generator with sodium chloride nuclei instead of fluorescein were performed. Results demonstrating applicability and limitations of DEHS particles for fluorometric deposition measurements are presented and discussed.

Growing concern about knowledge of aerosol transport in human lungs is caused by great potential of use of inhaled pharmaceuticals. Second substantial motive for the research is an effort to minimize adverse effects of particular matter emitted by traffic and industry on human health. We created model geometry of human lungs to 7th generation of branching. This model geometry was used for fabrication of two physical models. The first one is made from thin walled transparent silicone and it allows a measurement of velocity and size of aerosol particles by Phase Doppler Anemometry (PDA). The second one is fabricated by stereolithographic method and it is designed for aerosol deposition measurements. We provided a series of measurements of aerosol transport in the transparent model and we ascertained remarkable phenomena linked with lung flow. The results are presented in brief. To gather how this phenomena affects aerosol deposition in human lungs we used the second model and we developed a technique for deposition fraction and deposition efficiency assessment. The results confirmed that non-symmetric and complicated shape of human airways essentially affects transport and deposition of aerosol. The research will now focus on deeper insight in aerosol deposition.

The interaction between fluorescently labeled hyaluronan and cationic surfactants was studied using Fluorescence Correlation Spectroscopy. The hyaluronan was selected at two different molecular weights - specifically, 274 kDa and 710 kDa. Cetyltrimethylammonium bromide and Septonex® were chosen as cationic surfactants to interact with the negatively charged biopolymer. The study focused on changes in the diffusive behavior of a biopolymer that interacts with surfactant molecules in an aqueous environment. Various methods were applied to evaluate the obtained data, these including, among others, the Maximum Entropy Method, which provides the distributional dependences of diffusion coefficients. Without the surfactant, the studied biopolymers showed diffusion behavior comparable to that found in previously published studies. In the presence of surfactants, more intense interaction was observed between Cetyltrimethylammonium bromide and Septonex®. Comparing the molecular weights, the retention of intermolecular aggregates after the precipitation region for the lower weight and the disintegration of these aggregates for the higher weight were observed; moreover, they showed diffusion behavior comparable to the samples without the presence of the surfactant., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Liposomal carrier systems have emerged as a promising technology for pulmonary drug delivery. This study focuses on two selected liposomal systems, namely, dipalmitoylphosphatidylcholine stabilized by phosphatidic acid and cholesterol (DPPC-PA-Chol) and dipalmitoylphosphatidylcholine stabilized by polyethylene glycol and cholesterol (DPPC-PEG-Chol). First, the research investigates the stability of these liposomal systems during the atomization process using different kinds of nebulizers (air-jet, vibrating mesh, and ultrasonic). The study further explores the aerodynamic particle size distribution of the aerosol generated by the nebulizers. The nebulizer that demonstrated optimal stability and particle size was selected for more detailed investigation, including Andersen cascade impactor measurements, an assessment of the influence of flow rate and breathing profiles on aerosol particle size, and an in vitro deposition study on a realistic replica of the upper airways. The most suitable combination of a nebulizer and liposomal system was DPPC-PA-Chol nebulized by a Pari LC Sprint Star in terms of stability and particle size. The influence of the inspiration flow rate on the particle size was not very strong but was not negligible either (decrease of D v 50 by 1.34 μm with the flow rate increase from 8 to 60 L/min). A similar effect was observed for realistic transient inhalation. According to the in vitro deposition measurement, approximately 90% and 70% of the aerosol penetrated downstream of the trachea using the stationary flow rate and the realistic breathing profile, respectively. These data provide an image of the potential applicability of liposomal carrier systems for nebulizer therapy. Regional lung drug deposition is patient-specific; therefore, deposition results might vary for different airway geometries. However, deposition measurement with realistic boundary conditions (airway geometry, breathing profile) brings a more realistic image of the drug delivery by the selected technology. Our results show how much data from cascade impactor testing or estimates from the fine fraction concept differ from those of a more realistic case.

N,N,N-Trimethyl chitosan (TMC), a biocompatible and biodegradable derivative of chitosan, is currently used as a permeation enhancer to increase the translocation of drugs to the bloodstream in the lungs. This article discusses the effect of TMC on a mimetic pulmonary surfactant, Curosurf®, a low-viscosity lipid formulation administered to preterm infants with acute respiratory distress syndrome. Curosurf® exhibits a strong interaction with TMC, resulting in the formation of aggregates at electrostatic charge stoichiometry. At nanoscale, Curosurf® undergoes a profound reorganization of its lipid vesicles in terms of size and lamellarity. The initial micron-sized vesicles (average size 4.8 μm) give way to a froth-like network of unilamellar vesicles about 300 nm in size. Under such conditions, neutralization of the cationic charges by pulmonary surfactant may inhibit TMC permeation enhancer capacity, especially as electrostatic charge complexation is found at low TMC content. The permeation properties of pulmonary surfactant-neutralized TMC should then be evaluated for its applicability as a permeation enhancer for inhalation in the alveolar region., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

This contribution is focused on the preparation of a liposomal drug delivery system of erlotinib resisting the nebulization process that could be used for local treatment of non-small-cell lung cancer. Liposomes with different compositions were formulated to reveal their influence on the encapsulation efficiency of erlotinib. An encapsulation efficiency higher than 98 % was achieved for all vesicles containing phosphatidic acid (d ≈ 100 nm, ζ = - 43 mV) even in the presence of polyethylene glycol (d ≈ 150 nm, ζ = - 17 mV) which decreased this value in all other formulas. The three most promising formulations were nebulized by two air-jet and two vibrating mesh nebulizers, and the aerosol deposition in lungs was calculated by tools of computational fluid and particle mechanics. According to the numerical simulations and measurements of liposomal stability, air-jet nebulizers generated larger portion of the aerosol able to penetrate deeper into the lungs, but the delivery is likely to be more efficient when the formulation is administered by Aerogen Solo vibrating mesh nebulizer because of a higher portion of intact vesicles after the nebulization. The leakage of encapsulated drug from liposomes nebulized by this nebulizer was lower than 2 % for all chosen vesicles., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

UV-induced fingerprint spectroscopy (UV-IFS), a new tool in a toolbox of analytical methods, is a powerful technique registering molecule-specific changes of fluorescence induced by UV irradiation. Analysis of fluorescence spectra of a sample prior and after UV irradiation enables an identification of a sample of a drug or pharmaceutics based on a comparison with signals of known standards. Moreover, UV-IFS uncovers the presence of undesired contaminations or intentional changes of the composition. Herein, we employ UV-IFS for qualitative as well as quantitative analysis of common medicines including analgesic/antipyretic (Acetaminophen), antihistamines (Loratadine and Desloratadine), and phosphodiesterase type 5 inhibitors (Tadalafil and Sildenafil citrate). UV irradiation (λ em  = 254 nm) for 2 - 10 min induced significant changes of fluorescence of the studied samples and according to the unique patterns, the quality and quantity were evaluated. Limits of detection for individual active ingredients were calculated as follows: Acetaminophen = 0.1 µg·mL -1 , Loratadine = 0.1 μg·mL -1 , Desloratadine = 0.01 µg·mL -1 , Tadalafil = 0.04 µg·mL -1 and Sildenafil = 0.2 µg·mL -1 . Moreover, genuine and fake CIALIS, VIAGRA and KAMAGRA tablets were reliably identified., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Interactions of hyaluronan with micelles formed by cationic surfactants were studied by the time-resolved measurement of fluorescence resonance energy transfer (FRET) using perylene and fluorescein as probes. Two surfactants were studied - Cetyltrimethylammonium bromide (CTAB) and Septonex. In pure micellar solutions, the same values of FRET efficiency were found for both surfactants, but values for the binding of the probe pair were lower for Septonex micelles than in the case of CTAB. This was attributed to steric effects of the carbethoxy group in the Septonex polar head. Upon the addition of hyaluronan, decreased FRET efficiency and increased binding were detected in comparison with pure surfactants. To resolve the structure of the formed aggregates, steady state and time-resolved fluorescence anisotropy was employed as an additional technique. All results indicated that cationic micelles bind to hyaluronan forming a pearl necklace structure with micelles of smaller size compared to pure surfactant. Besides theoretical interest, the studied polyelectrolyte-surfactant system may be interesting for the formulation of drug delivery systems., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Cellular autofluorescence is usually considered to be a negative phenomenon because it can affect the sensitivity of fluorescence microscopic or flow cytometric assays by interfering with the signal of various fluorescent probes. Nevertheless, in our work, we adopted a different approach, and green autofluorescence induced by flavins was used as a tool to monitor fermentation employing the bacterium Cupriavidus necator . The autofluorescence was used to distinguish microbial cells from abiotic particles in flow cytometry assays, and it was also used for the determination of viability or metabolic characteristics of the microbial cells. The analyses using two complementary techniques, namely fluorescence microscopy and flow cytometry, are simple and do not require labor sample preparation. Flavins and their autofluorescence can also be used in a combination with other fluorophores when the need for multi-parametrical analyses arises, but it is wise to use dyes that do not emit a green light in order to not interfere with flavins' emission band (500-550 nm).

In this work, hydrogels formed by interaction of biopolymeric electrolytes and oppositely charged surfactants are studied from the point of view of their ability to incorporate model hydrophobic dyes in their micelle-like structure. Two types of hydrogels were investigated. The first type was based on cationized dextran cross-linked by sodium dodecylsulphate. The second type was prepared by interactions of hyaluronan with carbethoxypendecinium bromide (septonex). Nile red and Atto488 were used as model dyes for the diffusion experiments. The dyes were dissolved in two different media: surfactant and physiological saline. The diffusion of dyes into hydrogel was monitored over time. Effective diffusion coefficients were determined. It was found that their values are strongly influenced by the hydrogel character, the types of dye used and the solvent. The obtained effective coefficients were higher in comparison with the values determined for the diffusion in the opposite direction (release from the hydrogel). The dyes are presented as free in physiological saline and in the form of micelles or micelle aggregates in surfactants. During diffusion into the hydrogel, they can be gradually incorporated in a "pearl necklace structure" which suppresses their mobility. In contrast, this partial immobilization of dyes can increase the concentration gradient which is a driving force of diffusion. Also, the gradual incorporation of dyes into hydrogel structures influences the values of the effective diffusion coefficients., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Here, we present the potential analytical applications of photochemistry in combination with fluorescence fingerprinting. Our approach analyzes the fluorescence of samples after ultraviolet light (UV) treatment. Especially in presence of metal ions and thiol-containing compounds, the fluorescence behavior changes considerably. The UV-induced reactions (changes) are unique to a given sample composition, resulting in distinct patterns or fingerprints (typically in the 230-600 nm spectral region). This method works without the need for additional chemicals or fluorescent probes, only suitable diluent must be used. The proposed method (UV fingerprinting) suggests the option of recognizing various types of pharmaceuticals, beverages (juices and wines), and other samples within only a few minutes. In some studied samples (e.g. pharmaceuticals), significant changes in fluorescence characteristics (mainly fluorescence intensity) were observed. We believe that the fingerprinting technique can provide an innovative solution for analytical detection., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

In this study, three different molecules (cholesterol, phosphatidic acid, and polyethylene glycol) were used for the stabilization of liposomes during the nebulization process. The purpose of this article is to answer the question of whether the change in the composition of liposomes affected the parameters of generated aerosol and whether the nebulization process affected observed properties of liposomes. Firstly, liposomes with different composition were prepared and their properties were checked by dynamic and electrophoretic light scattering. The membrane properties were measured by fluorescence spectroscopy - especially generalized polarization (Laurdan) and anisotropy (Diphenylhexatriene). The same characteristic of liposomes was measured after the nebulization by vibrating mesh nebulizer. Cholesterol was capable of liposome stabilization because of increased membrane fluidity. The membrane properties of the outer and inner parts were not influenced by the nebulization process. Electrostatic stabilization was successful for the lowest concentration of phosphatidic acid, but after the nebulization process the hydration of the membrane outer part was changed. Higher amount of PEG needs to be added for successful steric stabilization. The nebulization process of the two lowest concentrations of PEG slightly influenced immobilized water and the rigidity of inner part of the membrane (especially around the phase transition temperature)., (Copyright © 2021 Elsevier B.V. All rights reserved.)

High-resolution ultrasound spectroscopy (HR-US), size and ζ-potential titrations, and isothermal titration calorimetry (ITC) were used to characterize the interactions between hyaluronan and catanionic ion pair amphiphile vesicles composed of hexadecyltrimethylammonium-dodecylsulphate (HTMA-DS), dioctadecyldimethylammonium chloride (DODAC), and cholesterol. In addition to these methods, visual observations were performed with the selected molecular weight of hyaluronan. A very good correlation was obtained between data from size titration, HR-US, and visual observation, which indicated in lower charge ratios the formation of hyaluronan-coated vesicles. On the contrary, at higher charge ratios, coated vesicles disintegrated to a size of around 2000 nm. The intensity of these interactions and the disaggregation were dependent on the molecular weight of hyaluronan. All interactions studied by ITC showed strong exothermic behavior, and these interactions between vesicles and hyaluronan were confirmed from the first addition, independently of the molecular weight of hyaluronan.

This work is focused on the study of the effect of cholesterol on the properties of vesicular membranes of ionic amphiphilic pairs at different temperatures. The hexadecyltrimethylammonium-dodecyl sulfate ionic amphiphilic pair system with the addition of 10 mol % dioctadecyldimethylammonium chloride was chosen for a detailed study of vesicle properties. A large range of cholesterol concentrations (0-73 mol %) in the temperature range 10-80 °C was studied. Under these conditions, the size distribution, the membrane fluidity, and the surface layer were monitored together with the change in the mobility of water in the surface layer. Obtained quantities were correlated with each other and combined into appropriate graphs. It was found that in stable systems that meet the condition of unimodal size distribution with a PDI value lower than 0.3, temperature has virtually no effect on the size of vesicular systems. On the contrary, when studying the hydration and fluidity of the membrane, significant changes in these parameters were found, which, however, do not affect the short-term stability of these vesicular systems. The presented results thus indicate the possibility of adjusting the composition of the vesicular system in terms of fluidity and membrane hydration while maintaining short-term stability and size distribution.

Herein, we report a new simple and easy-to-use approach for the characterization of protein oligomerization based on fluorescence resonance energy transfer (FRET) and capillary electrophoresis with LED-induced detection. The FRET pair consisted of quantum dots (QDs) used as an emission tunable donor (emission wavelength of 450 nm) and a cyanine dye (Cy3), providing optimal optical properties as an acceptor. Nonoxidative dimerization of mammalian metallothionein (MT) was investigated using the donor and acceptor covalently conjugated to MT. The main functions of MTs within an organism include the transport and storage of essential metal ions and detoxification of toxic ions. Upon storage under aerobic conditions, MTs form dimers (as well as higher oligomers), which may play an essential role as mediators in oxidoreduction signaling pathways. Due to metal bridging by Cd 2+ ions between molecules of metallothionein, the QDs and Cy3 were close enough, enabling a FRET signal. The FRET efficiency was calculated to be in the range of 11-77%. The formation of MT dimers in the presence of Cd 2+ ions was confirmed by MALDI-MS analyses. Finally, the process of oligomerization resulting in FRET was monitored by CE, and oligomerization of MT was confirmed., Competing Interests: Declaration of competing interest Authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Inorganic nanoparticles might have played a vital role in the transition from inorganic chemistry to self-sustaining living systems. Such transition may have been triggered or controlled by processes requiring not only versatile catalysts but also suitable reaction surfaces. Here, experimental results showing that multicolor quantum dots might have been able to participate as catalysts in several specific and nonspecific reactions, relevant to the prebiotic chemistry are demonstrated. A very fast and easy UV-induced formation of ZnCd quantum dots (QDs) with a quantum yield of up to 47% was shown to occur 5 min after UV exposure of the solution containing Zn(II) and Cd(II) in the presence of a thiol capping agent. In addition to QDs formation, xanthine activity was observed in the solution. The role of solar radiation to induce ZnCd QDs formation was replicated during a stratospheric balloon flight.

A capillary electrophoretic (CE) method using a short-sweep approach and laser-induced fluorescence (LIF) detection (ShortSweepCE-LIF) was developed for determination of Zn 2+ and Cd 2+ as complexes with highly selective and sensitive fluorescent probe FluoZin-3. The ShortSweepCE-LIF method, established in this work, can be used for examining competitive Zn 2+ and Cd 2+ binding properties of metalloproteins or peptides. The parameters including background electrolyte composition, injection pressure and time as well as separation voltage were investigated. Under the optimized conditions, 80 mM HEPES, pH 7.4, with 1.5 μM FluoZin-3 was used as an electrolyte, hydrodynamic injection was performed at 50 mbar for 5 s, and separation voltage of 25 kV. Limits of detection for Zn 2+ and Cd 2+ were 4 and 125 nM, respectively. The developed method was demonstrated in a study of interactions between metalothionein-2a isoform and metal ions Zn 2+ , Co 2+ and Cd 2+ . It was found that FluoZin-3 was able to extract a single Zn 2+ ion, while added Co 2+ (in surplus) extracted only 2.4 Zn 2+ ions, and Cd 2+ extracted all 7 Zn 2+ ions present in the metalothionein molecule., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The phase behavior of aqueous systems containing hyaluronan, at concentrations between 2 and 100 mg/L, and oppositely charged surfactants was investigated. A fluorescence probe technique revealed the formation of micellar structures on the hyaluronan in homogeneous systems well below the surfactant standard, critical, micellar concentration. Moreover, regions of gel-phase separation were revealed. A detailed phase diagram was, thus, constructed in the very diluted region and the hyaluronan concentration was found to be the main parameter controlling the phase behavior, in contrast to the charge ratio. The stability of hyaluronan-surfactant aggregates in the homogeneous systems while in storage at 4 °C (up to three months), against dilution, salt addition and on heating-cooling (between 10 and 50 °C) was also investigated. The aggregates were stable while in storage or upon increasing and decreasing the temperature. The dilution of hyaluronan-surfactant complexes or the addition of 0.15 M NaCl led to their disintegration. Finally, systems prepared in a 0.15 M NaCl solution showed that interactions are suppressed and no aggregation below the standard critical micellar concentration was observed., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This work explores molecular, morphological as well as biotechnological features of the highly promising polyhydroxyalkanoates (PHA) producer Halomonas halophila. Unlike many other halophiles, this bacterium does not require expensive complex media components and it is capable to accumulate high intracellular poly(3-hydroxybutyrate) (PHB) fractions up to 82% of cell dry mass. Most remarkably, regulating the concentration of NaCl apart from PHB yields influences also the polymer's molecular mass and polydispersity. The bacterium metabolizes various carbohydrates including sugars predominant in lignocelluloses and other inexpensive substrates. Therefore, the bacterium was employed for PHB production on hydrolysates of cheese whey, spent coffee grounds, sawdust and corn stover, which were hydrolyzed by HCl; required salinity of cultivation media was set up during neutralization by NaOH. The bacterium was capable to use all the tested hydrolysates as well as sugar beet molasses for PHB biosynthesis, indicating its potential for industrial PHB production., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Numerous prokaryotes accumulate polyhydroxyalkanoates (PHA) in the form of intracellular granules. The primary function of PHA is the storage of carbon and energy. Nevertheless, there are numerous reports that the presence of PHA granules in microbial cells enhances their stress resistance and fitness when exposed to various stress factors. In this work, we studied the protective mechanism of PHA granules against UV irradiation employing Cupriavidus necator as a model bacterial strain. The PHA-accumulating wild type strain showed substantially higher UV radiation resistance than the PHA non-accumulating mutant. Furthermore, the differences in UV-Vis radiation interactions with both cell types were studied using various spectroscopic approaches (turbidimetry, absorption spectroscopy, and nephelometry). Our results clearly demonstrate that intracellular PHA granules efficiently scatter UV radiation, which provides a substantial UV-protective effect for bacterial cells and, moreover, decreases the intracellular level of reactive oxygen species in UV-challenged cells. The protective properties of the PHA granules are enhanced by the fact that granules specifically bind to DNA, which in turn provides shield-like protection of DNA as the most UV-sensitive molecule. To conclude, the UV-protective action of PHA granules adds considerable value to their primary storage function, which can be beneficial in numerous environments.

Numerous prokaryotes accumulate polyhydroxybutyrate (PHB) intracellularly as a storage material. It has also been proposed that PHB accumulation improves bacterial stress resistance. Cupriavidus necator and its PHB non-accumulating mutant were employed to investigate the protective role of PHB under hypertonic conditions. The presence of PHB granules enhanced survival of the bacteria after exposure to hypertonic conditions. Surprisingly, when coping with such conditions, the bacteria did not utilize PHB to harvest carbon or energy, suggesting that, in the osmotic upshock of C. necator, the protective mechanism of PHB granules is not associated with their hydrolysis. The presence of PHB granules influenced the overall properties of the cells, since challenged PHB-free cells underwent massive plasmolysis accompanied by damage to the cell membrane and the leakage of cytoplasm content, while no such effects were observed in PHB containing bacteria. Moreover, PHB granules demonstrated "liquid-like" properties indicating that they can partially repair and stabilize cell membranes by plugging small gaps formed during plasmolysis. In addition, the level of dehydration and changes in intracellular pH in osmotically challenged cells were less pronounced for PHB-containing cultures, demonstrating the important role of PHB for bacterial survival under hyperosmotic conditions., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A freeze-drying method enabling solubilization of hydrophobic species in aqueous solutions of native hyaluronan is described. The method is based on opening the access to supposed hydrophobic patches on hyaluronan by disturbing its massive hydration shell. Hydrophobic and/or polarity-sensitive fluorescence probes were used as hydrophobic models or indicators of interactions with hydrophobic patches. Fluorescence parameters specific to individual probes confirmed the efficiency of the freeze-drying method. This work is the first step in developing biocompatible and biodegradable carriers for hydrophobic drugs with targeted distribution of the active compound from native, chemically non-modified hyaluronan.

A well-defined neutron spectrum is essential for many types of experimental topics and is also important for both calibration and testing of spectrometric and dosimetric detectors. Provided it is well described, such a spectrum can also be employed as a reference neutron field that is suitable for validating selected cross sections. The present paper aims to compare calculations and measurements of such a well-defined spectra in geometrically similar cores of the LR-0 reactor with fuel containing slightly different enrichments (2%, 3.3% and 3.6%). The common feature to all cores is a centrally located dry channel which can be used for the insertion of studied materials. The calculation of neutron and gamma spectra was realized with the MCNP6 code using ENDF/B-VII.0, JEFF-3.1, JENDL-3.3, ROSFOND-2010 and CENDL-3.1 nuclear data libraries. Only minor differences in neutron and gamma spectra were found in the comparison of the presented reactor cores with different fuel enrichments. One exception is the gamma spectrum in the higher energy region (above 8MeV), where more pronounced variations could be observed., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Accumulation of polyhydroxybutyrate (PHB) seems to be a common metabolic strategy adopted by many bacteria to cope with cold environments. This work aimed at evaluating and understanding the cryoprotective effect of PHB. At first a monomer of PHB, 3-hydroxybutyrate, was identified as a potent cryoprotectant capable of protecting model enzyme (lipase), yeast (Saccharomyces cerevisiae) and bacterial cells (Cupriavidus necator) against the adverse effects of freezing-thawing cycles. Further, the viability of the frozen-thawed PHB accumulating strain of C. necator was compared to that of the PHB non-accumulating mutant. The presence of PHB granules in cells was revealed to be a significant advantage during freezing. This might be attributed to the higher intracellular level of 3-hydroxybutyrate in PHB accumulating cells (due to the action of parallel PHB synthesis and degradation, the so-called PHB cycle), but the cryoprotective effect of PHB granules seems to be more complex. Since intracellular PHB granules retain highly flexible properties even at extremely low temperatures (observed by cryo-SEM), it can be expected that PHB granules protect cells against injury from extracellular ice. Finally, thermal analysis indicates that PHB-containing cells exhibit a higher rate of transmembrane water transport, which protects cells against the formation of intracellular ice which usually has fatal consequences.

Many bacteria are capable of accumulating intracellular granules of polyhydroxyalkanoates (PHA). In this work, we developed confocal microscopy analysis of bacterial cells to study changes in the diameters of cells as well as PHA granules during growth and PHA accumulation in the bacterium Cupriavidus necator H16 (formerly Ralstonia eutropha). The cell envelope was stained by DiD(®) fluorescent probe and PHA granules by Nile Red. Signals from both probes were separated based on their spectral and fluorescence life-time properties. During growth and PHA accumulation, bacterial cells increased their length but the width of the cells remained constant. The volume fraction of PHA granules in cells increased during PHA accumulation, nevertheless, its value did not exceed 40 vol. % regardless of the PHA weight content. It seems that bacterial cultures lengthen the cells in order to control the PHA volume portion. However, since similar changes in cell length were also observed in a PHA non-accumulating mutant, it seems that there is no direct control mechanism, which regulates the prolongation of the cells with respect to PHA granules volume. It is more likely that PHA biosynthesis and the length of cells are influenced by the same external stimuli such as nutrient limitation., (© FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Poly(3-hydroxybutyrate) (PHB) is a common carbon- and energy-storage compound simultaneously produced and degraded into its monomer 3-hydroxybutyrate (3HB) by numerous bacteria and Archae in a metabolic pathway called the PHB cycle. We investigated 3HB as a chemical chaperone capable of protecting model enzymes, namely lipase and lysozyme, from adverse effects of high temperature and oxidation. Heat-mediated denaturation of lipase in the presence or absence of 3HB was monitored by dynamic light scattering (DLS) revealing a significant protective effect of 3HB which increased as its concentration rose. Furthermore, when compared at the same molar concentration, 3HB showed a greater protective effect than the well-known chemical chaperones trehalose and hydroxyectoine. The higher protective effect of 3HB was also confirmed when employing differential scanning calorimetry (DSC) and lysozyme as a model enzyme. Furthermore, 3HB was capable of protecting lipase not only against thermal-mediated denaturation but also against oxidative damage by Cu(2+) and H2O2; its protection was higher than that of trehalose and comparable to that of hydroxyectoine. Taking into account that the PHB-producing strain Cupriavidus necator H16 reveals a 16.5-fold higher intracellular concentration than the PHB non-producing mutant C. necator PHB(-4), it might be expected that the functional PHB cycle might be responsible for maintaining a higher intracellular level of 3HB which, aside from other positive aspects of functional PHB metabolism, enhances stress resistance of bacterial strains capable of simultaneous PHB synthesis and mobilization. In addition, 3HB can be used in various applications and formulations as an efficient enzyme-stabilizing and enzyme-protecting additive.

Quaternary benzo[c]phenanthridine alkaloids are secondary metabolites of the plant families Papaveraceae, Rutaceae, and Ranunculaceae with anti-inflammatory, antifungal, antimicrobial and anticancer activities. Their spectral changes induced by the environment could be used to understand their interaction with biomolecules as well as for analytical purposes. Spectral shifts, quantum yield and changes in lifetime are presented for the free form of alkaloids in solvents of different polarity and for alkaloids bound to DNA. Quantum yields range from 0.098 to 0.345 for the alkanolamine form and are below 0.033 for the iminium form. Rise of fluorescence lifetimes (from 2-5 ns to 3-10 ns) and fluorescence intensity are observed after binding of the iminium form to the DNA for most studied alkaloids. The alkanolamine form does not bind to DNA. Acid-base equilibrium constant of macarpine is determined to be 8.2-8.3. Macarpine is found to have the highest increase of fluorescence upon DNA binding, even under unfavourable pH conditions. This is probably a result of its unique methoxy substitution at C12 a characteristic not shared with other studied alkaloids. Association constant for macarpine-DNA interaction is 700000 M(-1).

Viscosity-an integral property of a liquid-is traditionally determined by mechanical instruments. The most pronounced disadvantage of such an approach is the requirement of a large sample volume, which poses a serious obstacle, particularly in biology and biophysics when working with limited samples. Scaling down the required volume by means of microviscometry based on tracking the Brownian motion of particles can provide a reasonable alternative. In this paper, we report a simple microviscometric approach which can be conducted with common laboratory equipment. The core of this approach consists in a freely available standalone script to process particle trajectory data based on a Newtonian model. In our study, this setup allowed the sample to be scaled down to 10 μl. The utility of the approach was demonstrated using model solutions of glycerine, hyaluronate, and mouse blood plasma. Therefore, this microviscometric approach based on a newly developed freely available script can be suggested for determination of the viscosity of small biological samples (e.g., body fluids).