Search Results (14)

An optical sensor based on a polymethacrylate matrix (PMM) with immobilized gold nanoparticles (Au⁰ NPs) has been developed for the determination of pollutants in environmental samples. The nanoparticles are synthesized by chemical reduction of Au(III) to Au⁰ using sodium borohydride, which yields conglomerates of spherical particles with an absorption maximum at 530 nm. The time stability of the nanocomposite is demonstrated, as well as the ability to control the nanoparticle loading in the matrix by varying the concentration of the HAuCl₄ solution. The analytical capability of the PMM–Au⁰ system is demonstrated for the direct determination of tetracycline in river water in two linear concentration ranges: 0.001–0.010 mg/L and 0.025–0.100 mg/L, with detection limits of 0.0005 mg/L and 0.012 mg/L, respectively. The determination of tetracycline is based on the enhancement of its intrinsic fluorescence at 520 nm by gold nanoparticles in the solid phase following solid-phase extraction from water in the anionic form H₂TC⁻ using PMM–Au⁰. The colorimetric determination of thiocyanate anions is based on a color change of the PMM–Au⁰ nanocomposite from red to blue, corresponding to a shift in the plasmon absorption maximum from 530 nm to 630 nm. The sensor exhibits a linear response in the thiocyanate concentration range of 0.3–50.0 mg/L, with a detection limit of 0.1 mg/L. Thus, the multifunctional PMM–Au⁰ sensor has been used for the determination of various analytes employing different modes of analytical signal readout after minimal sample preparation. Full article

A polymethacrylate matrix (PMM) is proposed for the solid-phase extraction and determination of tetracycline (TC). The study of the influence of medium acidity, temperature, and contact time on the extraction of tetracycline by PMM showed that tetracycline is extracted by the matrix in the form of a singly charged anion H₂TC⁻, within the pH range of 8.9–9.7, with distribution coefficients reaching (5–6) × 10³ mL/g. Following the extraction process using PMM and PMM-Au⁰, the direct determination of tetracycline in the solid phase is possible without an elution step. This is achieved by using as the analytical signal both the intrinsic absorption and the instrumentally measured peak area of the anionic form of tetracycline, H₂TC⁻, in the matrix, with detection limits of 0.03 and 0.01 mg/L, respectively, and the fluorescence of tetracycline in PMM and PMM-Au⁰, with detection limits of 0.001 and 0.005 mg/L, respectively. The applicability of the digital colorimetry method for the quantitative determination of tetracycline based on its fluorescence in the solid phase is demonstrated. Methodologies for the determination of tetracycline using PMM and PMM-Au⁰ were developed and tested in the analysis of river and bottled water samples, biological fluid, as well as honey and milk samples. Full article

Organic semiconductor materials are interesting due to their application in various organic electronics devices. [1]benzothieno[3,2-b][1]benzothiophene (BTBT) is a widely used building block for the creation of such materials. In this work, three novel solution-processable regioisomeric derivatives of BTBT—2,7-bis(3-octylthiophene-2-yl)BTBT (1), 2,7-bis(4-octylthiophene-2-yl)BTBT (2), and 2,7-bis(5-octylthiophene-2-yl)BTBT (3)—were synthesized and investigated. Their optoelectronic properties were characterized experimentally by ultraviolet–visible and fluorescence spectroscopy, time-resolved fluorimetry, and cyclic voltammetry and studied theoretically by Time-Dependent Density Functional Theory calculations. Their thermal properties were investigated by a thermogravimetric analysis, differential scanning calorimetry, polarizing optical microscopy, and in situ small-/wide-angle X-ray scattering measurements. It was shown that the introduction of alkyl substituents at different positions (3, 4, or 5) of thiophene moieties attached to a BTBT fragment significantly influences the optoelectronic properties, thermal stability, and phase behavior of the materials. Thin films of each compound were obtained by drop-casting, spin-coating and doctor blade techniques and used as active layers for organic field-effect transistors. All the OFETs exhibited p-channel characteristics under ambient conditions, while compound 3 showed the best electrical performance with a charge carrier mobility up to 1.1 cm²·V⁻¹s⁻¹ and current on/off ratio above 10⁷. Full article

The discovery of new peptides and their derivatives is an outcome of ongoing efforts to identify a peptide with significant biological activity for effective usage as a possible therapeutic agent. Spinorphin peptides have been documented to exhibit numerous applications and features. In this study, biologically active peptide derivatives based on novel peptide analogues of spinorphin conjugated with 5,5′-dimethyl (Dm) and 5,5′-diphenyl (Ph) hydantoin derivatives have been successfully synthesized and characterized. Scanning electron microscopy (SEM) and spectral methods such as UV-Vis, FT-IR (Fourier Transform Infrared Spectroscopy), CD (Circular Dichroism), and fluorimetry were used to characterize the microstructure of the resulting compounds. The results revealed changes in peptide morphology as a result of the restructuring of the aminoacidic sequences and aromatic bonds, which is related to the formation of intermolecular hydrogen bonds between tyrosyl groups and the hydantoin moiety. Electrochemical and fluorescence approaches were used to determine some physicochemical parameters related to the biological behavior of the compounds. The biological properties of the spinorphin derivatives were evaluated in vivo for anticonvulsant activity against the psychomotor seizures at different doses of the studied peptides. Both spinorphin analog peptides with Ph and Dm groups showed activity against all three phases of the seizure in the intravenous Pentylenetetrazole Seizure (ivPTZ) test. This suggests that hydantoin residues do not play a crucial role in the structure of spinorphin compounds and in determining the potency to raise the seizure threshold. On the other hand, analogs with a phenytoin residue are active against the drug-resistant epilepsy test (6-Hz test). In addition, bioactivity analyses revealed that the new peptide analogues have the potential to be used as antimicrobial and antioxidant compounds. These findings suggest promising avenues for further research that may lead to the development of alternative medicines or applications in various fields beyond epilepsy treatment. Full article

Peptides and their related compounds can self-assemble into diverse nanostructures of different shapes and sizes in response to various stimuli such as pH, temperature or ionic strength. Here we report the synthesis and characterization of a lysozyme derived pentapeptide and its ability to build well-defined fibrillar structures. Lysozyme FESNF peptide fragment was synthesized by solid phase peptide synthesis using the Fmoc/t-Bu strategy, purified by analytical high-performance liquid chromatography (HPLC) and its molecular weight was confirmed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI–MS). Spectroscopic features of this pentapeptide were investigated by UV-visible spectroscopy and fluorimetry showing the pattern of marginal phenylalanine residues within the peptide sequence. Self-assembling properties were determined using atomic force microscopy (AFM), aggregation index and thioflavin T assay (ThT). FESNF generating fibrillar structures observed by AFM and aggregation propensity were primarily influenced by pH conditions. Moreover, the experimental data were confirmed by molecular dynamics simulation studies. The obtained fibrils will be used next to explore their potential to act as support material for medical and cosmetic application. Full article

For the first time, the pharmacokinetic (PK) profile of tryptophanol-derived isoindolinones, previously reported as p53 activators, was investigated. From the metabolites’ identification, performed by liquid chromatography coupled to high resolution tandem mass spectrometry (LC-HRMS/MS), followed by their preparation and structural elucidation, it was possible to identify that the indole C2 and C3 are the main target of the cytochrome P450 (CYP)-promoted oxidative metabolism in the tryptophanol-derived isoindolinone scaffold. Based on these findings, to search for novel p53 activators a series of 16 enantiopure tryptophanol-derived isoindolinones substituted with a bromine in indole C2 was prepared, in yields of 62–89%, and their antiproliferative activity evaluated in human colon adenocarcinoma HCT116 cell lines with and without p53. Structural optimization led to the identification of two (S)-tryptophanol-derived isoindolinones 3.9-fold and 1.9-fold more active than hit SLMP53-1, respectively. Compounds’ metabolic stability evaluation revealed that this substitution led to a metabolic switch, with the impact of Phase I oxidative metabolism being minimized. Through differential scanning fluorimetry (DSF) experiments, the most active compound of the series in cell assays led to an increase in the protein melting temperature (T_m) of 10.39 °C, suggesting an effective binding to wild-type p53 core domain. Full article

Plant sterols are known for their health-promoting effects, lowering blood cholesterol levels and alleviating cardiovascular disease. In this work, we continue our research on the asymmetric acylglycerols in which fatty acid residues are replaced by two stigmasterol residues in sn-1 and sn-2 or sn-2 and sn-3 positions as new thermostable carriers of phytosterols for their potential application in foods or as components of new liposomes in the pharmaceutical industry. The aim of this manuscript was to compare and analyze the effects of four distigmasterol-modified acylglycerols (dStigMAs) on the fluidity and the main phase transition temperature of the model phospholipid membrane. Their properties were determined using differential scanning calorimetry (DSC), steady-state fluorimetry and attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR). The determination of the effect of the tested compounds on the mentioned physicochemical parameters of the model membranes will allow for the determination of their properties and stability, which is essential for their practical application. The results indicated that all compounds effect on the physicochemical properties of the model membrane. The degree of these changes depends on the structure of the compound, especially the type of linker by which stigmasterol is attached to the glycerol backbone, as well as on the type of hydrocarbon chain. Full article

Henipaviruses are severe human pathogens within the Paramyxoviridae family. Beyond the P protein, the Henipavirus P gene also encodes the V and W proteins which share with P their N-terminal, intrinsically disordered domain (NTD) and possess a unique C-terminal domain. Henipavirus W proteins antagonize interferon (IFN) signaling through NTD-mediated binding to STAT1 and STAT4, and prevent type I IFN expression and production of chemokines. Structural and molecular information on Henipavirus W proteins is lacking. By combining various bioinformatic approaches, we herein show that the Henipaviruses W proteins are predicted to be prevalently disordered and yet to contain short order-prone segments. Using limited proteolysis, differential scanning fluorimetry, analytical size exclusion chromatography, far-UV circular dichroism and small-angle X-ray scattering, we experimentally confirmed their overall disordered nature. In addition, using Congo red and Thioflavin T binding assays and negative-staining transmission electron microscopy, we show that the W proteins phase separate to form amyloid-like fibrils. The present study provides an additional example, among the few reported so far, of a viral protein forming amyloid-like fibrils, therefore significantly contributing to enlarge our currently limited knowledge of viral amyloids. In light of the critical role of the Henipavirus W proteins in evading the host innate immune response and of the functional role of phase separation in biology, these studies provide a conceptual asset to further investigate the functional impact of the phase separation abilities of the W proteins. Full article

The aim of this work was the evaluation of the physico-chemical properties of a new type of liposomes that are composed of DPPC and bioconjugates of anisic acid with phosphatidylcholine. In particular, the impact of modified anisic acid phospholipids on the thermotropic parameters of liposomes was determined, which is crucial for using them as potential carriers of active substances in cancer therapies. Their properties were determined using three biophysical methods, namely differential scanning calorimetry (DSC), steady-state fluorimetry and attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR). Moreover, temperature studies of liposomes composed of DPPC and bioconjugates of anisic acid with phosphatidylcholine provided information about the phase transition, fluidity regarding chain order, hydration and dynamics. The DSC results show that the main phase transition peak for conjugates of anisic acid with phosphatidylcholine molecules was broadened and shifted to a lower temperature in a concentration- and structure-dependent manner. The ATR-FTIR results and the results of measurements conducted using fluorescent probes located at different regions in the lipid bilayer are in line with DSC. The results show that the new bioconjugates with phosphatidylcholine have a significant impact on the physico-chemical properties of a membrane and cause a decrease in the temperature of the main phase transition. The consequence of this is greater fluidity of the lipid bilayer. Full article

We aimed to characterize the mechanisms involved in neuroprotection by R-PIA administered before pilocarpine-induced seizures. Caspase-1 and caspase-3 activities were assayed using fluorimetry, and cathepsin D, HSP-70, and AKT expression levels were assayed using Western Blot of hippocampal samples. R-PIA was injected before pilocarpine (PILO), and four groups were studied at 1 h 30 min and 7 days following initiation of status epilepticus (SE): PILO, R-PIA+PILO, SALINE, and R-PIA+SALINE. At 1 h 30 min, significantly higher activities of caspase-1 and -3 were observed in the PILO group than in the SALINE group. Caspase-1 and -3 activities were higher in the R-PIA+PILO group than in the PILO group. At 7 days following SE, caspase-1 and -3 activities were higher than in the initial post-seizure phase compared to the SALINE group. The pretreatment of rats receiving PILO significantly reduced caspase activities compared to the PILO group. Expression of HSP-70, AKT, and cathepsin D was significantly higher in the PILO group than in the SALINE. In the R-PIA+PILO group, the expression of AKT and HSP-70 was greater than in rats receiving only PILO, while cathepsin D presented decreased expression. Pretreatment with R-PIA in PILO-injected rats strongly inhibited caspase-1 and caspase-3 activities and cathepsin D expression. It also increased expression levels of the neuroprotective proteins HSP-70 and AKT, suggesting an important role in modulating the cellular survival cascade. Full article

A well-known approach to the optical measure of oxygen is based on the quenching of luminescence by molecular oxygen. The main challenge for this measuring method is the determination of an accurate mathematical model for the sensor response. The reason is the dependence of the sensor signal from multiple parameters (like oxygen concentration and temperature), which are cross interfering in a sensor-specific way. The common solution is to measure the different parameters separately, for example, with different sensors. Then, an approximate model is developed where these effects are parametrized ad hoc. In this work, we describe a new approach for the development of a learning sensor with parallel inference that overcomes all these difficulties. With this approach we show how to generate automatically and autonomously a very large dataset of measurements and how to use it for the training of the proposed neural-network-based signal processing. Furthermore, we demonstrate how the sensor exploits the cross-sensitivity of multiple parameters to extract them from a single set of optical measurements without any a priori mathematical model with unprecedented accuracy. Finally, we propose a completely new metric to characterize the performance of neural-network-based sensors, the Error Limited Accuracy. In general, the methods described here are not limited to oxygen and temperature sensing. They can be similarly applied for the sensing with multiple luminophores, whenever the underlying mathematical model is not known or too complex. Full article

The classical approach to non-linear regression in physics is to take a mathematical model describing the functional dependence of the dependent variable from a set of independent variables, and then using non-linear fitting algorithms, extract the parameters used in the modeling. Particularly challenging are real systems, characterized by several additional influencing factors related to specific components, like electronics or optical parts. In such cases, to make the model reproduce the data, empirically determined terms are built in the models to compensate for the difficulty of modeling things that are, by construction, difficult to model. A new approach to solve this issue is to use neural networks, particularly feed-forward architectures with a sufficient number of hidden layers and an appropriate number of output neurons, each responsible for predicting the desired variables. Unfortunately, feed-forward neural networks (FFNNs) usually perform less efficiently when applied to multi-dimensional regression problems, that is when they are required to predict simultaneously multiple variables that depend from the input dataset in fundamentally different ways. To address this problem, we propose multi-task learning (MTL) architectures. These are characterized by multiple branches of task-specific layers, which have as input the output of a common set of layers. To demonstrate the power of this approach for multi-dimensional regression, the method is applied to luminescence sensing. Here, the MTL architecture allows predicting multiple parameters, the oxygen concentration and temperature, from a single set of measurements. Full article

Luminescence-based sensors for measuring oxygen concentration are widely used in both industry and research due to the practical advantages and sensitivity of this type of sensing. The measuring principle is the luminescence quenching by oxygen molecules, which results in a change of the luminescence decay time and intensity. In the classical approach, this change is related to an oxygen concentration using the Stern-Volmer equation. This equation, which in most cases is non-linear, is parameterized through device-specific constants. Therefore, to determine these parameters, every sensor needs to be precisely calibrated at one or more known concentrations. This study explored an entirely new artificial intelligence approach and demonstrated the feasibility of oxygen sensing through machine learning. The specifically developed neural network learns very efficiently to relate the input quantities to the oxygen concentration. The results show a mean deviation of the predicted from the measured concentration of 0.5% air, comparable to many commercial and low-cost sensors. Since the network was trained using synthetically generated data, the accuracy of the model predictions is limited by the ability of the generated data to describe the measured data, opening up future possibilities for significant improvement by using a large number of experimental measurements for training. The approach described in this work demonstrates the applicability of artificial intelligence to sensing technology and paves the road for the next generation of sensors. Full article

High spectral resolution spectrometers were used to detect optical signals ofongoing plant stress in potted white clover canopies subjected to ozone fumigation. Thecase of ozone stress is used in this manuscript as a paradigm of oxidative stress. Steadystatefluorescence (Fs) and the Photochemical Reflectance Index (PRI) were investigatedas advanced hyperspectral remote sensing techniques able to sense variations in the excessenergy dissipation pathways occurring when photosynthesis declines in plants exposed to astress agent. Fs and PRI were monitored in control and ozone fumigated canopies during a21-day experiment together with the traditional Normalized Difference Vegetation Index(NDVI) and physiological measurements commonly employed by physiologists to describestress development (i.e. net CO₂ assimilation, active fluorimetry, chlorophyll concentrationand visible injuries). It is shown that remote detection of an ongoing stress through Fs andPRI can be achieved in an early phase, characterized by the decline of photosynthesis. Onthe contrary, NDVI was able to detect the stress only when damage occurred. These resultsopen up new possibilities for assessment of plant stress by means of hyperspectral remotesensing. Full article